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| United States Patent |
5,741,799
|
|
Kimball
, et al.
|
April 21, 1998
|
Heterocyclic thrombin inhibitors
Abstract
Heterocyclic thrombin inhibitors are provided which have the structure
##STR1##
wherein n, R, R.sup.1, R.sup.2, R.sup.3, G, G.sub.x, R.sup.6', Ra, Xa,
R.sup.6, Rb, R.sub.3, p, Q, A and R.sup.4 are as defined herein.
| Inventors:
|
Kimball; Spencer D. (East Windsor, NJ);
Das; Jagabandhu (Mercerville, NJ);
Lau; Wan Fang (Lawrenceville, NJ);
Hall; Steven E. (Chapel Hill, NC);
Han; Wen-Ching (Newtown, PA)
|
| Assignee:
|
Bristol-Myers Squibb Company (Princeton, NJ)
|
| Appl. No.:
|
555561 |
| Filed:
|
November 8, 1995 |
| Current U.S. Class: |
514/316; 514/19; 514/210.18; 514/217.11; 514/218; 514/227.8; 514/237.2; 514/307; 514/311; 514/314; 514/315; 514/318; 514/319; 514/326; 514/343; 514/345; 514/374; 514/422; 514/423; 540/480; 540/481; 540/483; 540/544; 540/553; 540/575; 540/596; 540/597; 540/602; 540/603; 540/607; 540/610; 546/145; 546/146; 546/153; 546/187; 546/194; 546/206; 546/207; 546/208; 546/209; 546/210; 546/212; 546/214; 546/245; 546/279.1; 548/146; 548/212; 548/314.7; 548/517; 548/537; 548/538; 548/572 |
| Intern'l Class: |
A61K 031/445; A61K 031/40; C07D 401/12; C07D 207/09; C07D 211/28; 319 |
| Field of Search: |
514/211,212,227.8,237.2,314,252,218,316,318,326,365,374,422,423,343,315,307,311
540/480,481,483,544,553,575,597,602,603,610,596,607
544/60,129,141,360,372,130
546/206,207,208,209,210,212,214,279.1,216,245,146,153,145,165,194,187
548/517,572,212,146,314.7,537,548
|
References Cited
U.S. Patent Documents
| 4244865 | Jan., 1981 | Ali et al. | 514/326.
|
| 4258192 | Mar., 1981 | Okamoto et al. | 546/166.
|
| 4316889 | Feb., 1982 | Sandor et al. | 514/326.
|
| 4346078 | Aug., 1982 | Bajusz et al. | 514/326.
|
| 4904661 | Feb., 1990 | Pilgrim et al. | 514/237.
|
| 5002964 | Mar., 1991 | Lascalzo | 514/423.
|
| 5153176 | Oct., 1992 | Abe et al. | 514/18.
|
| 5518735 | May., 1996 | Sturzebecher | 424/449.
|
| Foreign Patent Documents |
| 479489 | Apr., 1992 | EP.
| |
| 526877 | Feb., 1993 | EP.
| |
| 0669317 | Mar., 1995 | EP.
| |
| WO931115 | Jun., 1993 | WO.
| |
| WO9429336 | Dec., 1994 | WO.
| |
Other References
Stuerzebecher J. Markwardt F, Walsmann P, Voigt B, Wagner G. Pharmazie,
42(2), 114-16, 1987.
Rubini E, Gilon C, Selinger Z, Chorer M. Tetrahedron 42(21), 6039-45, 1986.
Robert M. Knabb et al, "In Vivo Characterization of a New Synthetic
Thrombin Inhibitor," Thrombosis and Hemostasis (1992) 67, 56-59.
Charles V. Jackson et al, "Pharmacological Assessment of the Antithrombotic
Activity of the Peptide Thrombin Inhibitor,
D-Methyl-Phenylalanyl-Prolyl-Arginal (GYKI-14766), in a Canine Model of
Coronary Artery Thrombosis," J. Pharm. Exp. Ther. (1992) 261, 546-552.
Greissbach, U. et al "Peptide aldehydes as inhibitors of proteolytic
enzymes-synthetic aspects"CA102:181323a (1983).
Shuman, R.T. et al "Highly Selective Tripeptide Thrombin Inhibitors" J.
Med. Chem. 36, 314-19 (1993).
Bajusz, S. et al "Highly Active and Selective Anticoagulants" J. Med. Chem.
33, 1729-35 (1990).
Rubini, E. et al "synthesis of Isosteric Methylene-oxy Pseudodipeptide
Analogues as Novel Amide Bond Surrogate Units" Tetrahedron 42, 6039-45
(1986).
Spatola, A.F. et al "Amide Bond Surrogates: Pseudopeptides and Macrocycles"
Tetrahedron 44, 821-833 (1988).
McOmie, J.F.W. Protective Groups in Organic Chemistry Plenum Press, p. 46,
55-57, 73 (1973).
Banner, D. et al "Serine Proteases: 3D Structures, Mechanisms of Action and
Inhibitors"Prospect. Med. Chem. Bernard (Ed), Verlag Publishing, p. 27-43
(1993).
Hackh's Chemical Dictionary, McGraw-Hill, p. 16 (1982).
Burger, A. "A Guide to the Chemical Basis of Drug Design" Wiley, Science,
p. 15 (1984).
|
Primary Examiner: Ivy; C. Warren
Assistant Examiner: Huang; Evelyn
Attorney, Agent or Firm: Rodney; Burton
Parent Case Text
REFERENCE TO OTHER APPLICATIONS
This is a division of application Ser. No. 373,334, filed Jan. 17, 1995,
now U.S. Pat. No. 5,583,146 which is a continuation-in-part of application
Ser. No. 146,714 filed Nov. 10, 1993, now abandoned, which is a
continuation-in-part of application Ser. No. 112,153 filed Aug. 26, 1993,
now abandoned, which is a continuation-in-part of application Ser. No.
984,640 filed Dec. 2, 1992, now abandoned. Application Ser. No. 373,334 is
also a continuation-in-part of application Ser. No. 207,725 filed Mar. 14,
1994now abandoned; which is a continuation-in-part of application Ser. No.
56,279 filed May 3, 1993, now abandoned; and a continuation-in-part of
application Ser. No. 207,726 filed Mar. 14, 1994, now abandoned, which is
a continuation-in-part of application Ser. No. 56,017 filed May 3, 1993,
now abandoned, and a continuation-in-part of application Ser. No. 112,155
filed Aug. 26, 1993, now abandoned, and a continuation-in-part of
application Ser. No. 213,964 filed Mar. 16, 1994.
Claims
What is claimed is:
1. A compound having the structure Ia,
##STR234##
or a stereoisomer thereof, wherein n is 0, 1 or 2; R.sub.b is --A.sup.1
--R.sup.3a, --CO--A.sup.1 --R.sup.3a or --SO.sub.2 --A.sup.1 --R.sup.3a ;
wherein R.sup.3a is guanidine, amidine or amino, and A.sup.1 is an alkyl,
alkenyl or alkynyl chain of 2 to 6 carbons, provided that where R.sub.b is
--A.sup.1 --R.sup.3a, A.sup.1 is an alkenyl or alkynyl chain of 2 to 6
carbons; or
R.sub.b is --(CH.sub.2).sub.p --A.sup.2 --R.sup.2' or --(C.sub.2).sub.p
--CO--A.sup.2 R.sup.2' where p is 0, 1 or 2, R.sup.2' is amidine and
A.sup.2 is an azacycloalkyl, azaheteroalkyl or azaheteroalkenyl ring of 4
to 8 atoms, optionally substituted by alkyl, CO or halo as given by the
structure:
##STR235##
where X' is CH.sub.2, O, S or NH; p is 0, 1 or 2;
q=0, 1, 2, 3 or 4 if X'.dbd.CH.sub.2 ;
q=2,3 or 4 if X'.dbd.O, S, NH; and
y.sup.1, y.sup.2 are independently H, alkyl or halo; or
R.sub.b is --(CH.sub.2).sub.p --A.sup.3 --R.sup.4, --(CH.sub.2).sub.p
--CO--A.sup.3 --R.sup.4, or --(CH.sub.2).sub.p --SO.sub.2 --A.sup.3
--R.sup.4,
wherein R.sup.4 is guanidine, amidine or aminomethyl, and A.sup.3 is aryl
or cycloalkyl;
R is hydrogen, hydroxyalkyl, aminoalkyl, alkyl, cycloalkyl, aryl,
arylalkyl, alkenyl, alkynyl, amidoalkyl, arylalkoxyalkyl or an amino acid
side chain, either protected or unprotected;
R.sup.1 and R.sup.2 are independently hydrogen, lower alkyl, cycloalkyl,
aryl, hydroxy, alkoxy, oxo, thioxo, thioalkyl, thioaryl, amino or
alkylamino;
R.sup.6 is hydrogen,
##STR236##
--SO.sub.2 R.sup.7' or --CO.sub.2 R.sup.7' (wherein R.sup.7' is lower
alkyl, aryl or cyclohetero-alkyl);
or a pharmaceutically acceptable salt thereof.
2. A method of inhibiting or preventing formation of blood clots, which
comprises administering to a patient in need of treatment a antithrombin
effective amount of a compound as defined in claim 1.
3. A pharmaceutical composition comprising a antithrombin effective amount
of a compound as defined in claim 1 and a pharmaceutically acceptable
carrier therefor.
4. The compound as defined in claim 1 having the structure Ia,
##STR237##
or a stereoisomer thereof, wherein n is 0, 1 or 2; R.sub.b is
--(CH.sub.2).sub.p --A.sup.2 --A.sup.2' or --(CH.sub.2).sub.p
--CO--A.sup.2 R.sup.2' where p is 0, 1 or 2, R.sup.2' is amidine and
A.sup.2 is an azacycloalkyl, azaheteroalkyl or azaheteroalkenyl ring of 4
to 8 atoms, optionally substituted by alkyl, CO or halo as given by the
structure:
##STR238##
where X' is CH.sub.2, O, S or NH; p is 0, 1 or 2;
q=0, 1, 2, 3 or 4 if X'.dbd.CH.sub.2 ;
q=2,3 or 4 if X'.dbd.O, S, NH; and
y.sup.1, y.sup.2 are independently H, alkyl, or halo;
R.sup.1 and R.sup.2 are independently hydrogen, lower alkyl, cycloalkyl,
aryl, hydroxy, alkoxy, keto, thioketal, thioalkyl, thioaryl, amino or
alkylamino;
R is hydrogen, hydroxyalkyl, aminoalkyl, alkyl, cycloalkyl, aryl,
arylalkyl, alkenyl, alkynyl, amidoalkyl, arylalkoxyalkyl or an amino acid
side chain, either protected or unprotected; and
R.sup.6 is hydrogen,
##STR239##
--SO.sub.2 R.sup.7' or --CO.sub.2 R.sup.7' (wherein R.sup.7' is lower
alkyl, aryl or cyclohetero-alkyl);
or a pharmaceutically acceptable salt thereof.
5. The compound as defined in claim 4 wherein n is O, R.sup.6 is
2-naphthylsulfonyl, H, methyl-sulfonyl, benzyloxycarbonyl or
t-butoxycarbonyl, R is arylalkyl, hydroxyalkyl or benzyloxyalkyl, R.sup.1
is H, R.sup.2 is H and R.sub.b is
##STR240##
6. The compound as defined in claim 1 wherein R.sub.b is --A.sup.1
--R.sup.3a,
##STR241##
or --SO.sub.2 --A.sup.1 --R.sup.3 a.
7. The compound as defined in claim 1 wherein R.sub.b is --(CH.sub.2).sub.p
--A.sup.2 --R.sup.2' or
##STR242##
8. The compound as defined in claim 1 wherein R.sub.b is --(CH.sub.2).sub.p
--A.sup.3 --R.sup.4, --(CH.sub.2).sub.p --CO--A.sup.3 --R.sup.4 or
--(CH.sub.2).sub.p --SO.sub.2 --A.sup.3 --R.sup.4.
9. The compound as defined in claim 1 wherein n is O.
10. The compound as defined in claim 1 wherein R.sup.6 is
2-naphthylsulfonyl, H, benzyloxycarbonyl, t-butoxycarbonyl or
methylsulfonyl.
11. The compound as defined in claim 1 wherein R.sup.3a is guanidine or
amino.
12. The compound as defined in claim 1 having the structure
##STR243##
13. The compound as defined in claim 1 which is S-(R*
R*)!-4-(aminoiminomethyl)amino!-N-1-3-hydroxy-2-(2-naphthalenylsulfon
yl)amino!-1-oxoprop-yl!-2-pyrrolidinyl!-METHYL!butanamide, or a
pharmaceutically acceptable salt thereof;
R-(R*,S*)!-4-(aminoiminomethyl)amino!-N-1-(2-amino-1-oxo-3-phenylpropyl
)-2-pyrrolidinyl!-methyl!butanesulfonamide, or a pharmaceutically
acceptable salt thereof;
R-(R*,S*)!-2-2-3-(aminoiminomethyl)-amino!propyl!sulfonyl!amino!met
hyl!-1-pyrrolidinyl-2-oxo-1-(phenylmethyl)ethyl!carbamic acid,
1,1-dimethylethyl ester or a pharmaceutically acceptable salt thereof;
R-(R*,S*)!-3-(aminoiminomethyl)amino!-N-1-(2-amino-1-oxo-3-phenylpropyl
)-2-pyrrolidinyl!-methyl!propanesulfonamide, or a pharmaceutically
acceptable salts thereof;
R-(R*,S*)!-2-amino-1-2-4-(aminomethyl)-phenyl!-amino!METHYL!-1-pyrroli
dinyl!-3-phenyl-1-propanone, or a pharmaceutically acceptable salt thereof;
R-(R*,S*)!-N-2-2-4-(aminomethyl)phenyl!amino!METHYL!-1-pyrrolidinyl!-
2-oxo-1-(phenylmethyl)-ethyl!-2-naphthalenesulfonamide, or a
pharmaceutically acceptable salt thereof;
S-(R*,S*)!-4-(aminoiminomethyl)amino!-N-1-2-(methylsulfonyl)amino!-1-
oxo-3-phenylpropyl!-2-pyrrolidinyl!methyl!butanamide, or a pharmaceutically
acceptable salt thereof;
R-(R*,S*)!-2-amino-1-2-4-(aminomethyl)-phenyl!amino!METHYL!-1-pyrrolid
inyl!-3-phenyl-1-propanone, or a pharmaceutically acceptable salt thereof;
R-(R*,S*)!-N-2-2-4-(aminomethyl)phenyl!amino!METHYL!-1-pyrrolidinyl!-
2-oxo-1-(phenylmethyl)ethyl!-2-naphthalenesulfonamide, or a
pharmaceutically acceptable salt thereof;
S-(R*,R*)!-4-(aminoiminomethyl)amino!-N-1-3-hydroxy-2-(7-methoxy-2-n
aphthalenyl)sulfonyl!amino!-1-oxopropyl!-2-pyrrolidinyl!methyl!butanamide,
or a pharmaceutically acceptable salt thereof;
R-(R*,S*)!-N-2-2-4-(aminoiminomethyl)-phenyl!amino!METHYL!pyrrolidiny
l!-2-oxo-1-(phenylmethyl)ethyl!methanesulfonamide, or a pharmaceutically
acceptable salt thereof;
S-(R*,S*)!-4-1-(2-amino-1-oxo-3-phenyl-propyl)-2-pyrrolidinyl!METHYL!am
ino!benzenecarbox-imidamide, or a pharmaceutically acceptable salt thereof;
R-(R*,S*)!-N-2-2-4-(aminoiminomethyl)phenyl!amino!methyl!pyrrolidinyl
!-2-oxo-1-(phenylmethyl)ethyl!methanesulfonamide, or a pharmaceutically
acceptable salt thereof;
1(S),2S!-1-(aminoiminomethyl)-N-1-3-hydroxy-2-(2-naphthalenylsulfonyl)
amino!-1-oxo-propyl!-2-pyrrolidinyl!methyl!-3-piperidinecarboxamide, or a
pharmaceutically acceptable salt thereof;
S-(R*,R*)!-1-(aminoiminomethyl)-N-1-3-hydroxy-2-(2-naphthalenylsulfony
l)amino!-1-oxopropyl!-2-pyrrolidinyl!methyl!-4-piperidinecarboxamide, or a
pharmaceutically acceptable salt thereof;
N-(S)-2-(S)-2-1-(aminoiminomethyl)-3-piperidinyl!acetyl!amino!methyl!
-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide, or
a pharmaceutically acceptable salt thereof;
S-(R*,R*)-N-2-2-1-(aminoiminomethyl)-4-piperidinyl!acetyl!amino!meth
yl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
or a pharmaceutically acceptable salt;
S
(R*,R*)!-N-2-2-5-(aminoiminomethyl)amino!-1-oxopentyl!amino!methyl!-
1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonarnide, or
a pharmaceutically acceptable salt thereof;
S-(R*,R*)!-N-2-2-6-(aminoiminomethyl)amino!-1-oxohexyl!amino!methyl!
-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide, or
a pharmaceutically acceptable salt thereof;
1S2R*(3S*)!!-N-2-2-1-(aminoimino-methyl)-3-piperidinyl!carbonyl!ami
no!methl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfon
amide, or a pharmaceutically acceptable salt thereof;
S-(R*,R*)!-N-2-2-7-(aminoiminomethyl)amino!-1-oxoheptyl!amino!methyl
!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
or a pharmaceutically acceptable salt thereof;
1S2R*(3R*)!!-N-2-2-1-aminoimino-methyl)-3-piperidinyl!carbonyl!amin
o!methyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfon
amide, acceptable salt thereof;
S-(R*,R*)!-4-(aminomethyl)-N-1-3-hydroxy-2-(2-naphthalenylsulfonyl)ami
no!-1-oxopropyl!-2-pyrrolidinyl!methyl!benzamide, or a pharmaceutically
acceptable salt thereof;
1S(2R*,3R*)!-N-2-1-(aminoiminomethyl)-3-piperidinyl!carbonyl!amino!me
thyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-.alpha.-toluenesulfonam
ide or a pharmaceutically acceptable salt thereof:
1S(2R*,3R*)!-N-2-1-(aminoiminomethyl)-3-piperidinyl!carbonyl!amino!me
thyl!-1-pyrrolidinyl!-1-(carbomethoxymethyl)-2-oxoethyl-.alpha.-toluenesulf
onamide or a pharmaceutically acceptable salt thereof;
1S(2R*,3R*)!-N-2-1-(aminoiminomethyl)-3-piperidinyl!carbonyl!amino!me
thyl!-1-pyrrolidinyl!-1-(carboxamidomethyl)-2-oxoethyl!-.alpha.-toluenesulf
onamide or a pharmaceutically acceptable salt thereof;
S-(R*,R*)!-3-(aminomethyl)-N-1-3-hydroxy-2-(2-napthalenylsulfonyl)amino
!-1-oxopropyl!-2-pyrrolidinyl!methyl!benzamide, or a pharmaceutically
acceptable salt thereof;
S-(R*,R*)!-4-(aminomethyl)-N-1-3-hydroxy-2-(2-napthalenylsulfonyl)amino
!-1-oxopropyl!-2-pyrrolidinyl!methyl!benzeneacetamide, or a
pharmaceutically acceptable salt thereof;
S-(R*,R*)!-3-(aminomethyl)-N-1-3-hydroxy-2-(2-napthalenylsulfonyl)amin
o!-1-oxopropyl!-2-pyrrolidinyl!methyl!benzeneacetamide, or a
pharmaceutically acceptable salt thereof;
3S-3R*,3(R*,R*)!!-1-(aminoiminomethyl)-N-1-2-(2-napthalenylsulfonyl)a
mino!-1-oxopropyl1!-2-pyrrolidinyl!methyl!-3-piperidinecarboxamide, or a
pharmaceutically acceptable salt thereof;
3S-3R*,3(R*,R*)!!-1-(aminoiminomethyl)-N-1-3-hydroxy-2-(4-methylphen
yl)sulfonyl!amino!-1-oxopropyl!-2-pyrrolidinyl!METHYL!-3-piperidinecarboxam
ide, or a pharmaceutically acceptable salt thereof:
3S-3R*,3(R*,R*)!!-1-(aminoiminomethyl)-N-1-3-hydroxy-2-carbobenzylox
y!amino!-1-oxopropyl!-2-pyrrolidinyl!methyl!-3-piperidinecarboxamide, or a
pharmaceutically acceptable salt thereof;
1S2(R*,S*)!!-N-2-1-aminoiminomethyl)-4-piperidinyl!carbonyl!amino!me
thyl!-1-piperidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide
, or a pharmaceutically acceptable salt thereof;
S-(R*,R*)!-3-(aminoiminomethyl)amino!-N-1-3-hydroxy-2-2-naphthalenes
ulfonyl!amino!-1-oxopropyl!-2-pyrrolidinyl!methyl!propionamide, or a
pharmaceutically acceptable salt thereof;
1S2(R*,S*),(3R*)!!-N-2-1-(aminoimino-methyl)-3-piperidinyl!carbonyl!
amino!methyl!-1-piperidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesul
fonamide, or a pharmaceutically acceptable salt thereof;
1S2R*!!-N-2-1-(aminoiminomethyl)-3-phenyl!carbonyl!amino!methyl!-1-p
yrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide, or a
pharmaceutically acceptable salt thereof;
1S2R*!!-N-2-1-(aminoiminomethyl)-cis-5-methyl-3-piperidinyl!carbonyl
!amino!methyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenes
ulfonamide, or a pharmaceutically acceptable salt thereof;
1S2R*!!-N-2-1-(aminoiminomethyl)-trans-5-methyl-3-piperidinyl!carbon
yl!amino!methyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalen
esulfonamide, or a pharmaceutically acceptable salt thereof;
1S(2R*,3R*,4R*)!-N-2-1-(aminoiminomethyl)-3-piperidinyl!carbonyl!amin
o!methyl!-1-pyrrolidinyl!-1-(1-hydroxyethyl)-2-oxoethyl!-2-naphthalenesulfo
namide, or a pharmaceutically acceptable salt;
1S(2R*,3S*,4R*)!-N-2-1-(aminoiminomethyl)-3-piperidinyl!carbonyl!amino
!methyl!-1-pyrrolidinyl!-1-(1-hydroxyethyl)-2-oxoethyl!-2-naphthalenesulfon
amide, or a pharmaceutically acceptable salt thereof;
R-(S*,R*)!-N-2-2-1-(aminoiminomethyl)-4-piperidinyl!carbonyl!amino!m
ethyl!-1-pyrrolidinyl!-1-(phenylmethyl)-2-oxoethyl!methanesulfonamide, or a
pharmaceutically acceptable salt thereof;
S-(R*,R*)!-N-2-1-(aminoiminomethyl)-3-piperidinyl!carbonyl!amino!meth
yl-1-pyrrolidinyl!-2-oxoethyl!-2-naphthalenesulfonamide, or a
pharmaceutically acceptable salt thereof;
R-(S*,R*)!-N-2-2-1-(aminoiminomethyl)-4-piperidinyl!carbonyl!amino!m
ethyl!-1-pyrrolidinyl!-1-(methyl)-2-oxoethyl!benzylsulfonamide, or a
pharmaceutically acceptable salt thereof;
##STR244##
or a pharmaceutically acceptable salt thereof;
##STR245##
or a pharmaceutically acceptable salt thereof;
##STR246##
or a pharmaceutically acceptable salt thereof;
##STR247##
or a pharmaceutically acceptable salt thereof;
##STR248##
or a pharmaceutically acceptable salt thereof;
##STR249##
or a pharmaceutically acceptable salt thereof.
14. A compound according to claim 13 wherein the pharmaceutically
acceptable salt is a trifluoroacetate.
Description
FIELD OF THE INVENTION
The present invention relates to heterocyclic compounds which are thrombin
inhibitors and thus useful in inhibiting formation of thrombi.
DESCRIPTION OF THE FIRST EMBODIMENT OF THE INVENTION
In a first aspect or embodiment, the present invention relates to
sulfonamido heterocyclic thrombin inhibitors which have the structure I
##STR2##
wherein G is an amido moiety which is
##STR3##
including all stereoisomers thereof; and including all pharmaceutically
acceptable salts thereof; wherein
R is hydrogen, hydroxyalkyl, aminoalkyl, amidoalkyl, alkyl, cycloalkyl,
aryl, arylalkyl, alkenyl, alkynyl, arylalkoxyalkyl, or an amino acid side
chain, either protected or unprotected;
R.sup.1 and R.sup.2 are independently hydrogen, lower alkyl, cycloalkyl,
aryl, hydroxy, alkoxy, oxo (also referred to as keto), thioxo (also
referred to as thioketal), thioalkyl, thioaryl, amino or alkylamino; or
R.sup.1 and R.sup.2 together with the carbons to which they are attached
form a cycloalkyl, aryl, or heteroaryl ring; and
R.sup.3 is lower alkyl, aryl, arylalkyl, heteroaryl, quinolinyl or
tetrahydroquinolinyl;
n is 0, 1 or 2;
m is 0, 1, 2 or 3;
Y is NH or S;
p is 0, 1, or 2;
Q is a single bond or
##STR4##
A is aryl or cycloalkyl, or an azacycloalkyl ring A of 3 to 7 carbons in
the ring (4 to 8 total ring members) or an azaheteroalkyl ring A of 4 to 6
carbons in the ring (4 to 8 total ring members),
##STR5##
where X is CH.sub.2, O, S or NH;
q is 0, 1, 2, 3 or 4 if X is CH.sub.2 ;
q is 2, 3 or 4 if X is O, S or NH;
Y.sup.1 and Y.sup.2 are independently H, lower alkyl or halo; and
R.sup.4 is guanidine, amidine or aminomethyl;
where A is aryl or cycloalkyl, R.sup.4 is guanidine, amidine or
aminomethyl;
where A is azacycloalkyl or azaheteroalkyl, R.sup.4 is amidine;
provided that where X is a hetero atom (that is, A is azaheteroalkyl), then
there must be at least a 2-carbon chain between X and any N atom in the
ring A or outside ring A;
and provided that where G is G1, then if R.sup.3 is alkyl, the alkyl must
contain at least 3 carbons.
In another embodiment of the invention, in the formula I compounds, R.sup.3
is 10-camphor
##STR6##
In still another embodiment of the formula I compounds, R.sup.3 is
pentaalkylphenyl, or trialkylphenyl such as pentafluorophenyl,
pentamethylphenyl or 2,4,6-tri-isopropylphenyl.
In yet another embodiment of the formula I compounds, R.sup.3 is
3-carboxyphenyl, 3-trifluoromethylphenyl or 4-carboxyphenyl.
Examples of the A ring (azacycloalkyl or azaheteroalkyl) which may be
employed herein include
##STR7##
and the like.
Preferred are compounds of formula I wherein G is
##STR8##
wherein Q is a single bond and A is an azacycloalkyl ring
##STR9##
where q is 0 or 1; and
R.sup.4 is amidino;
R.sup.3 is lower alkyl or aryl;
R is aralkyl or hydroxyalkyl;
R.sup.1 and R.sup.2 are each H;
n is 0 or 1.
A more preferred embodiment of the heterocyclic thrombin inhibitors of the
invention has the structure IA
##STR10##
where R is aralkyl (preferably benzyl), aryl (preferably phenyl), or
arylalkoxyalkyl (preferably benzyloxymethyl) and alkyl is preferably
methyl, ethyl or propyl, including all stereoisomers thereof.
Other preferred compounds of formula I are those wherein G is Gl, n is 0 or
1; m is 2; R.sup.3 is aryl or alkyl; R is arylalkyl or hydroxyalkyl such
as hydroxymethyl; R.sup.1 is hydrogen or lower alkyl such as methyl or
ethyl; R.sup.2 is H; and Y is --NH--; and compounds of formula IB:
##STR11##
The compounds of formula I of the invention (first embodiment) wherein G is
##STR12##
and Y is NH may be prepared according to the following Reaction Sequence
I.
##STR13##
The compounds of formula I of the invention wherein G is
##STR14##
and Y is NH may also be prepared according to the following Reaction
Sequence II
##STR15##
As seen in the above Reaction Sequence I, compounds of formula I wherein Y
is --NH--, are prepared as follows. The ester II is made to undergo a
carbodiimide coupling reaction with protected amino acid III in the
presence of ethyl 3-(3-dimethylamino)propyl carbodiimide hydrochloride
(WSC) or dicyclohexylcarbodiimide (DCC), and 1-hydroxybehzotriazole
monohydrate (HOBT), and N-methylmorpholine (NMM), and in the presence of
an inert organic solvent such as dimethylformamide (DMF), THF or
N-methylpyrrolidone, to form the amide IV. Amide IV is deprotected by
treatment with trifluoroacetic acid with or without the presence of dry
inert organic solvent such as dichloromethane, chloroform or THF at
temperatures within the range of from about -15.degree. to about
20.degree. C. Sulfonyl chloride V is added followed by organic base such
as triethylamine, pyridine or N,N-diisopropylethylamine to form the
sulfonamide VI. Sulfonamide VI is hydrolyzed by treatment with alkali
metal base such as NaOH or LiOH in the presence of an alcohol solvent such
as methanol or ethanol. The reaction mixture is acidified with HCl,
KHSO.sub.4 or H.sub.2 SO.sub.4, to form acid VII. The acid VII is then
subjected to a carbodiimide coupling reaction wherein VII is treated with
protected amine VIII in the presence of WSC or DCC, and HOBT, and NMM, in
the presence of an inert organic solvent such as dimethylformamide, THF or
N-methylpyrrolidone, to form sulfonamide IX. The sulfonamide IX is then
dissolved in an alcohol solvent such as ethanol or methanol, to which HCl
has been added and the mixture is hydrogenated over Pd--C or Pd(OH).sub.2
--C in the case where P.sup.1 is carbobenzyloxy. The crude material is
separated by conventional procedures and the desired isomers are treated
with amidine sulfonic acid X in the presence of an alcohol solvent such as
ethanol to form the compound of the invention IB.
In a preferred preparation, compounds of formula IB can be prepared from a
compound of formula IX by deprotection of P.sup.1 and reaction with
1-(1'-carbobenzyloxy)carboxamidine pyrazole
##STR16##
to give the protected guanidine form of IB. The CBZ protecting group can
then be removed by hydrogenation.
Compounds of formula IB can also be prepared from a compound of formula VII
and a compound of formula
##STR17##
by reaction with a coupling agent such as WSC or DCC and HOBT and a
suitable base such as NMM or triethylamine. Hydrogenation over palladium
on carbon to remove the CBZ protecting group then affords IB. Further,
compounds of formula IB can be prepared by the direct reaction of a
compound of formula
##STR18##
with a coupling reagent such as WSC or DCC and a base such as NMM or
triethylamine to give IB.
As seen in the above Reaction Sequence II, compounds of formula I wherein Y
is --NH--, are prepared as follows. The protected acid IIA is made to
undergo a carbodiimide coupling reaction with protected amino acid VIII in
the presence of ethyl 3-(3-dimethylamino)propyl carbodiimide hydrochloride
(WSC) or dicyclohexylcarbodiimide (DCC), and 1-hydroxybenzotriazole
monohydrate (HOBT), and N-methylmorpholine (NMM), and in the presence of
an inert organic solvent such as dimethylformamide (DMF), THF or
N-methylpyrrolidone, to form the amide IXA. Amide IXA is deprotected by
treatment with trifluoroacetic acid (TFA) when P is t-butoxycarbonyl (BOC)
or H.sub.2 --Pd/C when P is carbobenzyloxy (CBz), with or without the
presence of dry inert organic solvent such as dichloromethane, chloroform
or THF, at temperatures within the range of from about -15.degree. to
about 20.degree. C. to form amide IXB. The amide IXB is then subjected to
a carbodiimide coupling reaction wherein IXB is treated with protected
amine III in the presence of WSC or DCC, and HOBT, and NMM, in the
presence of an inert organic solvent such as dimethylformamide, THF or
N-methylpyrrolidone, to form amide IXC. The amide IXC is then dissolved in
an alcohol solvent such as ethanol or methanol, to which HCl has been
added and the mixture is hydrogenated over Pd--C or Pd(OH).sub.2 --C in
the case where P.sup.1 is CBz or treated with trifluoroacetic acid when
P.sup.1 is BOC. The crude material is separated by conventional procedures
and the desired isomers are treated with amidine sulfonic acid X in the
presence of an alcohol solvent such as ethanol to form IXD. Compound IXD
is then deprotected by treatment with TFA when P is BOC or by treatment
with H.sub.2 -Pd/C when P is CBz, as described above, and sulfonyl
chloride V is added followed by organic base such as triethylamine,
pyridine or N,N-diisopropylethylamine to form the sulfonamide IB.
The compounds of formula I of the invention wherein G is
##STR19##
and Y is S may be prepared according to the following Reaction Sequence
III.
##STR20##
Referring to the above Reaction Sequence III, compounds of formula I
wherein Y=S can be prepared as follows. The acid VII is subjected to a
carbodiimide coupling reaction wherein VII is treated with an aminoalcohol
XI in the presence of WSC or DCC, HOBT, and NMM, in the presence of an
inert organic solvent such as dimethylformamide, THF or
N-methylpyrrolidone, to form sulfonamide alcohol XII. The sulfonamide
alcohol XII is reacted with p-toluenesulfonyl chloride (TsCl) in pyridine,
or in a solvent such as methylene chloride or chloroform, with
N,N-dimethylaminopyridine to provide toluenesulfonate XIII. The compound
IC (Y=S) is prepared by treating XIII with thiourea in a solvent such as
DMF or DMSO at temperatures within the range of from about 25.degree. C.
to about 100.degree. C.
The compounds of formulae I and IA of the invention wherein G is
##STR21##
wherein A is azacycloalkyl or azaheteroalkyl, and R.sup.4 is amidine, may
be prepared according to the following Reaction Sequence IV:
##STR22##
As seen in the above Reaction Sequence IV, compounds of formula I wherein G
is
##STR23##
and A is azacycloalkyl or azaheteroalkyl, are prepared as follows. The
protected acid XV is made to undergo a carbodiimide coupling reaction with
amine XVI in the presence of ethyl 3-(3-dimethylamino)propyl carbodiimide
hydrochloride (WSC) or dicyclohexylcarbodiimide (DCC), and
1-hydroxybenzotriazole monohydrate (HOBT), and N-methylmorpholine (NMM),
and in the presence of an inert organic solvent such as dimethylformamide
(DMF), THF or N-methylpyrrolidone, to form the amide XVII. Amide XVII is
deprotected by treatment with, for example, H.sub.2/ Pd--C, if P.sup.1 is
CBz, to form amine XVIII. Amine XVIII is treated with amidinesulfonic acid
X in the presence of alcohol solvent, such as ethanol to form amine XIX.
Amine XIX is deprotected by treatment with trifluoroacetic acid (if
P=BOC), with or without the presence of dry inert organic solvent such as
dichloromethane, chloroform or THF, at temperatures within the range of
from about -15.degree. to about 20.degree. C. Sulfonyl chloride V is added
followed by organic base such as triethylamine, pyridine or
N,N-diisopropylethylamine to form the sulfonamide ID of the invention.
Alternatively, the order of steps in reaction sequence IV may be changed.
Thus the doubly protected compound of formula XVII is subjected to
conditions which remove the protecting group P.sup.1 (e.g.,
trifluoroacetic acid in methylene chloride if P.sup.1 is BOC), after which
the free amino group is made to undergo a reaction with
bis-t-butoxycarbonyl thiourea as described in the literature (Tet. Lett.,
1992, 5933-5936). The product is then subjected to conditions that remove
protecting group P (e.g., hydrogenation over Pd--C if P is CBZ),
sulfonylated as described, and the BOC protecting groups removed (by e.g.,
trifluoroacetic acid) to give the compounds of formula ID.
In a preferred embodiment, compounds of formula ID can be prepared from
compounds of formula XVII by removal of the P protecting group, followed
by sulfonamide formation using R.sup.3 SO.sub.2 Cl. The P.sup.1 protecting
group of the resulting sulfonamide is then removed, and the product
reacted with, 1-(1-carbobenzyloxy)-carboxamidine pyrazole
##STR24##
to give the CBZ-protected guanidine. The CBZ group may be removed by
hydrogenation to give the guanidine.
Compounds of formula ID can also be prepared from a compound of formula VII
and a compound of formula
##STR25##
by reaction with WSC or DCC and HOBT and a suitable base such as NMM or
triethylamine. Hydrogenation over palladium on carbon to remove the CBZ
protecting group then affords ID. Further, compounds of formula ID can be
prepared by the direct reaction of compound of formula
##STR26##
with a coupling reagent such as WSC or DCC and a base such as NMM or
triethylamine to give ID.
The starting materials of formula XVI are known in the art or may be
prepared by those skilled in the art employing conventional techniques.
The compounds of formulae I and IA of the invention where G is
##STR27##
where A is aryl or cycloalkyl and R.sup.4 is amidine or guanidine may be
prepared according to the following Reaction Sequence V:
##STR28##
As seen in Reaction Sequence V, compounds of formulae I and IA where G is
##STR29##
are prepared as follows. The protected acid XV is subjected to a
carbodiimide coupling reaction wherein XV is treated with protected amine
XVIA in the presence of WSC or DCC, and HOBT, and NMM, in the presence of
an inert organic solvent such as dimethylformamide, THF or
N-methylpyrrolidone, to form amide XVIIA. The amide XVIIA is then
dissolved in an alcohol solvent such as ethanol or methanol, to which HCl
has been added and the mixture is hydrogenated over Pd--C or Pd(OH).sub.2
--C in the case where P is carbobenzyloxy. The amide XVIIIA is treated
with sulfonyl chloride V followed by base to form the compound of the
invention IE.
The starting compound XVIA is known in the art or may be prepared employing
conventional procedures.
The compounds of formulae I and IA of the invention wherein G is
##STR30##
where A is aryl or cycloalkyl (that is A.sup.1) and R.sup.4 is aminomethyl
(that is R.sup.4") may be prepared according to the following Reaction
Sequence VI:
##STR31##
As seen in the above Reaction Sequence VI, compounds of formulae I and IA
wherein G is
##STR32##
are prepared as follows. The protected acid XV is made to undergo a
carbodiimide coupling reaction with protected amino acid XVIB in the
presence of ethyl 3-(3-dimethylamino)propyl carbodiimide hydrochloride
(WSC) or dicyclohexylcarbodiimide (DCC), and 1-hydroxybenzotriazole
monohydrate (HOBT), and N-methylmorpholine (NMM), and in the presence of
an inert organic solvent such as dimethylformamide (DMF), THF or
N-methylpyrrolidone, to form the amide XVIIB. Amide XVIIB is deprotected
by treatment with trifluoroacetic acid (TFA) when P is t-butoxycarbonyl
(BOC) or H.sub.2 --Pd/C when P is carbobenzyloxy (CBz), with or without
the presence of dry inert organic solvent such as dichloromethane,
chloroform or THF, at temperatures within the range of from about
-15.degree. to about 20.degree. C. to form amide XVIIIB. The amide XVIIIB
is then subjected to a sulfonation reaction wherein amide XVIIIB is
reacted with sulfonyl chloride V in the presence of organic base such as
triethylamine, pyridine or N,N-diisopropylethylamine to form the
sulfonamide IF of the invention.
The starting compounds XVIB are known in the art or may be prepared
employing conventional procedures.
The starting acid XV may be prepared from ester IV by hydrolyzing ester IV
by treating with a base such as NaOH, KOH or LiOH and then neutralizing
the resulting alkali metal salt with strong acid such as HCl or oxalic
acid.
DESCRIPTION OF THE SECOND EMBODIMENT OF THE INVENTION
In a second aspect or embodiment, the present invention relates to
guanidinyl- or amidinyl-substituted heterocyclic thrombin inhibitors which
have the structure 1. 1.
##STR33##
including all stereoisomers thereof, wherein n is 0, 1 or 2; Xa is S, SO,
SO.sub.2 or O;
R.sub.a is --A.sub.1 --R.sup.3a, where A.sub.1 is an alkyl, alkenyl, or
alkynyl chain of 2 to 6 carbon atoms and R.sup.3a is guanidine, amidine,
or amino; or
R.sub.a is --(CH.sub.2).sub.p --A.sub.2 --R.sup.2' where R.sup.2' is
amidine and A.sub.2 is an azacycloalkyl ring of 4 to 8 atoms, optionally
substituted by alkyl or halo as given by the structure
##STR34##
where p is 0, 1 or 2, q is 0, 1, 2, 3 or 4, N and X.sub.a are separated by
at least two carbon atoms; and
Y.sup.1, Y.sup.2 are independently H, alkyl, or halo; or
R.sub.a is --(CH.sub.2).sub.p --A.sub.3 --R.sup.4,
wherein R.sup.4 is guanidine, amidine or aminomethyl, and A.sub.3 is aryl
or cycloalkyl;
R, R.sup.1 and R.sup.2 are as defined hereinbefore; and
R.sup.6' is hydrogen,
##STR35##
--SO.sub.2 R.sup.7 or --CO.sub.2 R.sup.7 (wherein R.sup.7 is lower alkyl,
aryl, cycloheteroalkyl or heteroaryl);
including pharmaceutically acceptable salts thereof, and all stereoisomers
thereof.
Preferred are compounds of formula 1. wherein Xa is S or SO.sub.2, n is 0,
R.sub.a is --(CH.sub.2).sub.p --A.sub.2 R.sup.2' or --CH.sub.2
(CH.sub.2).sub.z --R.sup.3a, z is 1, 2, 3 or 4; more preferably
##STR36##
or R.sub.a is
##STR37##
p is 1; q is 1 or 2, R.sup.1 and R.sup.2 are each H, R is H or --CH.sub.2
OH, and R.sup.6' is
##STR38##
The compounds of formula 1. of the invention (second embodiment) wherein Xa
is S, SO, SO.sub.2 or O may be prepared according to the following
reaction sequences.
##STR39##
Referring to Reaction Sequence 1. compounds of formula 1. wherein Xa is S
and R.sub.a is --A.sub.1 --R.sup.3a and R.sup.3a is amino may be prepared
starting with tosylate 2. (prepared as described in J. Med. Chem. 35, 2615
(1992)) (wherein PG in 2. is a protecting group such as
##STR40##
(fluorenylmethoxycarbonyl, FMOC), preferably BOC, which is subjected to a
displacement reaction wherein 2. is treated with potassium thioacetate
(KSAc) in the presence of an inert organic solvent such as acetone,
dimethylformamide (DMF) or tetrahydrofuran (THF), under an inert
atmosphere such as argon, at a temperature within the range of from about
0.degree. to about 100.degree. C. to form thioacetate 3. Thioacetate 3. is
then alkylated by reacting 3. with an alkali metal alkoxide such as sodium
methoxide or potassium t-butoxide in an inert organic solvent such as THF,
DMF or diethylether, in the presence of an alcohol solvent such as
methanol or ethanol under an inert atmosphere such as argon, at a
temperature within the range of from about -30.degree. to about 50.degree.
C. To the resulting solution is added N-bromoalkyl-phthalimide 3.sup.a to
form the thiophthalimide 4. Thiophthalimide 4. is deprotected by treatment
with a deprotecting agent such as trifluoroacetic acid where P.G. is BOC
or HBr/acetic acid where P.G. is CBZ, with or without the presence of dry
inert organic solvent such as dichloromethane, chloroform or THF, to form
a crude amine salt 4'.
##STR41##
The resulting salt 4' is made to undergo a carbodiimide coupling reaction
with protected amino acid 4.sup.a.
##STR42##
(wherein PG' is a protecting group which may be any of the PG protecting
groups) in the presence of 1-hydroxybenzotriazole monohydrate (HOBT) ,
ethyl-3-(3-dimethylamino)propyl carbodiimide (WSC) or
dicyclohexylcarbodiimide (DCC), and N-methylmorpholine (NMM), in the
presence of an inert organic solvent such as dimethylformamide (DMF), THF
or N-methylpyrrolidone, to form carbamate 5.
Carbamate 5. is deprotected by treatment with trifluoroacetic acid (TFA) or
other deprotecting agent such as HBr/HOAc (depending on P.G.'), with or
without the presence of dry inert organic solvent such as dichloromethane,
chloroform or THF, to give a crude trifluoroacetic acid amine salt 6.
(where the deprotecting agent is TFA).
The crude amine salt 6. is then subjected to a coupling reaction wherein it
is reacted with
a) R.sup.7 COOH in the presence of WSC or DCC, HOBT and NMM,
b) R.sup.7 SO.sub.2 Cl in the presence of triethylamine or
c) R.sup.7 OCOCl in the presence of triethylamine, to form 7.
7. is then reacted with anhydrous hydrazine in the presence of dry solvent
such as dichloromethane, chloroform or THF, and an alcohol solvent such as
methanol or ethanol, to form thioalkylamine compound 1.A of the invention.
Compounds of formula 1. of the invention where R.sup.6' is H, Xa is S and
R.sub.a is --A.sub.1 --R.sup.3a and R.sup.3a is NH.sub.2 may be prepared
as shown in Reaction Sequence 1.B) where amine 6. is reacted with
anhydrous hydrazine (as described above in the reaction of 7. ) to form
1.A'.
Compounds of formula 1. of the invention wherein Xa is S, R.sub.a is
--A.sub.1 -guanidinyl and R.sup.6' is not H may be prepared as shown in
Reaction Sequence 7.A) wherein 1.A is subjected to a guanylation by
reacting 1.A with amidine sulfonic acid or other guanylating agent such as
1H-pyrazole-1-carboxamidine, in the presence of a weak organic base such
as triethylamine, pyridine or N-methylmorpholine (NMM) and an alcohol
solvent such as ethanol or methanol, to form 1.B.
Compounds of formula 1. of the invention wherein Xa is S, R.sub.a is
--A.sub.1 -guanidinyl and R.sup.6 is H may be prepared as outlined in
Reaction Sequence 2.B wherein compound 5. is treated with hydrazine (as
described above in Sequence 1.A in the reaction of 6.) to form amine 5.A
which is subjected to a guanylation reaction (as described above in
Sequence 2.A) to form protected guanidine 5.B which is deprotected by
treatment with TFA where P.G.' is BOC or using other deprotecting agent as
described hereinbefore, to form compound of the invention 1.B'.
Referring to Reaction Sequence 3.A, compounds of formula 1. of the
invention wherein Xa is S and R.sub.a is --A.sub.1 -amidinyl and R.sup.6'
=H are prepared by alkylating 3. by treating 3. with an alkali metal
alkoxide and with nitrile (3b) in the presence of methanol or
dimethylformamide, at a temperature within the range of from about
-60.degree. to about 75.degree. C., to form nitrile 8. which is made to
undergo amidine formation by treating nitrile 8. with hydrochloric acid in
the presence of an alcohol such as methanol or ethanol, and then with
ammonia to form amidine compound 9. Compound 9. is then subjected to a
carbodiimide coupling reaction by treating 9. with (4.sup.a) in the
presence of WSC and HOBT (as described above in the reaction of 4. to form
5. in Sequence 1.), to form 10. which is deprotected as described
hereinbefore to form 1.C'.
Compounds of formula 1. of the invention wherein Xa is S, R.sub.a is
--A.sub.1 -amidinyl and R.sup.6' is not H, are prepared as shown in
Sequence 3.B where 1.C' is treated with a coupling agent R.sup.7 COOH,
R.sup.7 OCOCl or R.sup.7 SO.sub.2 Cl (as described above in Sequence 1.A)
in the coupling of 6. to form 7.), to thereby form 1.C.
Referring to Reaction Sequence 4., compounds of formula 1. wherein Xa is SO
and R.sub.a is --A.sub.1 --R.sup.3a and R.sup.3a is NH.sub.2, may be
prepared by oxidizing amide 6. or 7. by treating 6. or 7. with an
oxidizing agent such as m-chloroperbenzoic acid (MCPBA), oxone or sodium
periodate in the presence of an inert organic solvent such as
dichloromethane, chloroform or acetonitrile, employing from about 0.9 to
about 1.5 moles of oxidizing agent per mole of 6. or 7., to form the
corresponding sulfoxide (Xa is SO). The corresponding sulfone (Xa is
SO.sub.2) is formed employing from about 2 to about 3 moles of oxidizing
agent per mole of 6. or 7. The so-formed sulfoxide or sulfone is then
deprotected with hydrazine (as described in Sequence 1.A in the reaction
of 6.) to form the corresponding amine 1.D of the invention.
As seen in Sequence 4 B1 where R.sup.6 .noteq.H and R.sup.3a is guanidine,
the resulting amine 1.D is guanylated by reaction with amidine sulfonic
acid (as described hereinbefore in forming compounds of formula I where Xa
is S) to form compounds of formula 1. where Xa is SO or SO.sub.2 (that is
1.E).
Where R.sup.6' is H and R.sub.a is --A.sub.1 -guanidinyl (Sequence 4.B2)
compounds of formula 1. of the invention wherein Xa is SO or SO.sub.2 are
formed as follows: compound 5. is oxidized as described above for 6. or
7., to form compound 12. which is then reacted with hydrazine to form 13.
which is guanylated and then deprotected to form compound of the invention
1.E' employing procedures described hereinbefore.
Sulfoxides or sulfones of compounds of formula 1. of the invention where
R.sub.a is --A.sub.1 -amidinyl are prepared as shown in Sequence 4.C) by
oxidizing 1.C or 1.C' employing procedures as described above to form
compound of the invention 1.F.
Referring to Reaction Sequence 5., compounds of formula 1. wherein Xa is S
and R.sub.a is --A.sub.2 R.sup.2' and R.sup.2' is amidinyl, may be
prepared starting with tosylate 2. which is subjected to a displacement
reaction wherein 2. is treated with nitrile 2.sup.a
2.sup.a HS--A.sub.2 --CN
in the presence of a base such as sodium methoxide, triethylamine or
pyridine, to form nitrile 15. Nitrile 15. is admixed with strong acid such
as hydrochloric acid, and an alcohol solvent such as methanol, or ethanol
and made to undergo amidine formation by treatment with ammonia to form
the corresponding amidine 16. The resulting amidine compound 16. is then
subjected to a carbodiimide coupling reaction with protected amino acid
4.sup.a in the presence of WSC or DCC, and HOBT and NMM, as described with
respect to Sequence 1., to form amide 17. Amide 17. is deprotected by
treatment with trifluoroacetic acid or other deprotecting agent, as
described with respect to Sequence 1., to form compound 1.G of the
invention, which may be subjected to a coupling reaction wherein it is
reacted with R.sup.7 COOH, R.sup.7 SO.sub.2 Cl or R.sup.7 OCOCl, as
described with respect to Sequence 1., to form compounds of formula 1.H
wherein Xa is S and R.sub.a is --A.sub.2 R.sup.2' and R.sup.2' is
amidinyl.
Referring to Reaction Sequence 6., compounds of formula 1. of the invention
wherein Xa is S, R.sub.a is --A.sub.2 --R.sup.2' and R.sup.2' is
aminomethyl, may be prepared by reducing compound 15., for example using a
reducing agent such as lithium aluminum hydride, triethylborohydride or
diborane, to form amine 18. which is treated with a protecting agent
(18.sup.a) such as CBZ--Cl to form 19. Protected amine 19. is then treated
with HCl in the presence of an alcohol such as methanol or dioxane and
then treated with coupling agent (4.sup.a), as described hereinbefore with
respect to Sequence 1., to form compound 20. Compound 20. may be
completely deprotected with e.g., HBr/HOAc to give amine 1.J.
Alternatively, 20. may be selectively deprotected with e.g., TFA (if
PG'=BOC) and then subjected to a coupling reaction by treatment with
coupling agent R.sup.7 COOH, R.sup.7 OCOCl or R.sup.7 SO.sub.2 Cl,
followed by removal of PG.sup.2 with HBr/HOAc, to form amine 1.K of the
invention.
Referring to Reaction Sequence 7., compounds of formula 1. wherein Xa is S,
R.sub.a is --A.sub.2 --R.sup.2' and R.sup.2' is guanidinyl may be prepared
by guanylating amine 21. to form guanidine 22. which is reacted with 2. to
form guanidine 23. Guanidine 23. is deprotected and then made to undergo a
coupling reaction with 4.sup.a, as described with respect to Sequence 1.,
to form 24 which is deprotected to form guanidine 1.L of the invention.
Guanidine 1.L may then be reacted with a coupling agent R.sup.7 COOH,
R.sup.7 OCOCl or R.sup.7 SO.sub.2 Cl, as described with respect to
Sequence 1., to form guanidine 1.M of the invention.
Referring to Reaction Sequence 8., compounds of formula 1. of the invention
wherein R.sub.a is --A.sub.2 --R.sup.2' and Xa is SO may be prepared by
oxidizing 1.G, 1.H, 1.J, 1.K, 1.L or 1.M by treating same with an
oxidizing agent such as m-chloroperbenzoic acid (MCPBA), oxone or sodium
periodate in the presence of an inert organic solvent such as
dichloromethane, employing from about 0.9 to about 1.5 moles of oxidizing
agent per mole of amide, to form the corresponding sulfoxide. The
corresponding sulfone may be formed employing from about 2 to about 3
moles of oxidizing agent per mole of amide. The so-formed sulfoxide or
sulfone is then deprotected with hydrazine and the resulting amine is
guanylated by reaction with amidine sulfonic acid (as described
hereinbefore in forming compounds of formula 1. where Xa is S) to form
compounds of formula 1. where Xa is SO or SO.sub.2.
As seen in Reaction Sequence 9., compounds of formula 1.Q and 1.R may be
prepared by treating thioacetate 3. with sodium methoxide, followed by an
alkylating agent of formula 25. to provide a compound of formula 26.
Deprotection of 26. (by TFA if PG=BOC) and carbodiimide coupling, as
described previously, affords compound 27. Removal of the protecting group
PG' (by HBr/HOAc if PG'=CBZ) followed by guanylation using
1H-pyrazole-1-carboxamidine (JOC 1992, 57, 2497-2502) then provides a
compound of formula 1.Q where R.sup.6' is not H. The compound 1.Q can be
deprotected by piperidine (if R.sup.6' =fluorenylmethyloxycarbonyl (FMOC))
to obtain compounds 1.R where R.sup.6' =H.
The key step for the preparation of compounds wherein Xa is oxygen is shown
in Scheme 10. Treatment of the protected alcohol 28. with KOH and a
mesylate (Ms) of formula 29. in xylene (see J. Med. Chem 1986, 29,
2335-2347) will provide the series of compounds 30. Further steps in the
preparation of compounds of formula 1. wherein Xa is oxygen may be
effected in analogy with the previously described Schemes 1, 2, 3, 5, 6,
7, and 9.
DESCRIPTION OF THE THIRD EMBODIMENT OF THE INVENTION
In a third aspect or embodiment, the present invention relates to
guanidinyl- or amidinyl-substituted methylamino heterocyclic thrombin
inhibitors which have the structure Ia Ia.
##STR43##
including all stereoisomers thereof wherein n is 0, 1 or 2; R.sub.b is
--A.sup.1 --R.sup.3a, --CO--A.sup.1 --R.sup.3a or --SO.sub.2 --A.sup.1
--R.sup.3a ; wherein R.sup.3a is guanidine, amidine or amino, and A.sup.1
is an alkyl, alkenyl or alkynyl chain of 2 to 6 carbons; or
R.sub.b is --(CH.sub.2).sub.p --A.sup.2 --R.sup.2' or --(CH.sub.2).sub.p
--CO--A.sup.2 --R.sup.2' where p is 0, 1 or 2, R.sup.2' is amidine and
A.sup.2 is an azacycloalkyl, azaheteroalkyl or azaheteroalkenyl ring of 4
to 8 atoms, optionally substituted by alkyl, CO or halo as given by the
structure:
##STR44##
where X' is CH.sub.2, O, S or NH;
q=0, 1, 2, 3 or 4 if X'=CH.sub.2 ;
q=2, 3 or 4 if X'=O, S, NH; and
Y.sup.1, Y.sup.2 are independently H, alkyl, or halo; or
R.sub.b is --(CH.sub.2).sub.p --A.sup.3 --R.sup.4, --(CH.sub.2).sub.p
--CO--A.sup.3 --R.sup.4, or --(CH.sub.2).sub.p --SO.sub.2 --A.sup.3
--R.sup.4,
wherein R.sup.4 is guanidine, amidine or aminomethyl, A.sup.3 is aryl or
cycloalkyl, and p is as defined above;
R, R.sup.1 and R.sup.2 are as defined hereinbefore;
R.sup.6 is hydrogen,
##STR45##
--SO.sub.2 R.sup.7' or --CO.sub.2 R.sup.7' (wherein R.sup.7' is lower
alkyl, aryl or cycloheteroalkyl);
including pharmaceutically acceptable salts thereof.
Preferred are compounds of formula Ia wherein n is 0, R.sub.b is
##STR46##
or --CH.sub.2 (CH.sub.2).sub.q -guanidine, and q is 3, 4 or 5; and
compounds of formula Ia wherein R.sub.b is A.sup.3 --R.sup.4, A.sup.3 is
phenyl, and R.sup.4 is amidine and compounds of formula I.sup.a wherein
R.sub.b is --(CH.sub.2).sub.p --A.sup.2 --R.sup.2' or
##STR47##
p is 0 or 1, A.sup.2 is azacycloalkyl or azacycloalkenyl, and R.sup.2' is
amidine;
R.sup.1 and R.sup.2 are each H, R is hydroxymethyl, --CH.sub.2 COOalkyl, or
benzyl and R.sup.6 is
##STR48##
H, BOC or CBZ.
Most preferred are compounds of formula Ia wherein n is 0, R.sub.b is
--(CH.sub.2).sub.p --A.sup.2 --R.sup.2' or
##STR49##
wherein p is 0 or 1, A.sup.2 is
##STR50##
R.sup.2' is amidine, R.sup.1 and R.sup.2 are each H, R is hydroxymethyl,
--CH.sub.2 COOCH.sub.3, or benzyl, and R.sup.6 is
##STR51##
H, BOC or CBZ.
The compounds of formula Ia of the invention (third embodiment) may be
prepared according to the following reaction sequences.
##STR52##
As seen in Reaction Sequence A, Part A, compounds of formula Ia of the
invention wherein R.sub.b is
##STR53##
and R.sup.3a is guanidine are prepared as follows. Tosylate IIa (prepared
as described by J. Das et al, J. Med. Chem., 1992, Vol. 35, 2610) (wherein
P.G. represents a protecting group such as t-butyloxy carbonyl (BOC),
carbobenzyloxy (CBZ), or fluorenyl-methyloxycarbonyl (FMOC) is made to
undergo a displacement reaction with sodium azide in the presence of an
inert organic solvent such as dimethylsulfoxide (DMSO) or
dimethylformamide (DMF), at a temperature within the range of from about
50.degree. to about 90.degree. C., to form azide IIIa which is reduced by
reaction with a reducing agent such as H.sub.2 /Pd--C or
triphenylphospine/H.sub.2 O, lithium aluminum hydride (LAH) or stannous
chloride, to form the corresponding amine IVa.
The amine IVa is made to undergo a carbodiimide coupling reaction with
protected amino acid IV' (where PG' is a protecting group such as any of
the PG protecting groups set out above) in the presence of ethyl
3-(3-dimethylamino)propyl carbodiimide hydrochloride (WSC) or
dicyclohexylcarbodiimide (DCC), and 1-hydroxybenzotriazole monohydrate
(HOBT), and N-methylmorpholine (NMM), in the presence of an inert organic
solvent such as dimethylformamide (DMF), THF or N-methylpyrrolidone (NMP),
to form the amide Va. Amide Va is deprotected by treatment with
trifluoroacetic acid (TFA) (where PG is BOC) or with H.sub.2 /Pd--C (where
PG is CBZ) with or without the presence of dry inert organic solvent such
as dichloromethane, chloroform or THF, at temperatures within the range of
from about -15.degree. to about 20.degree. C., to form amide VIa. Amide
VIa is then made to undergo a carbodiimide coupling reaction with
protected amino acid VI' in the presence of WSC or DCC, HOBT and NMM, as
described above with respect to amine IVa, to form compound VIIa. The
protecting group PG of VIIa is removed by reacting VIIa with TFA where PG
is BOC and PG' is other than BOC, to form partially deprotected compound
VIIIa. Compound VIIIa is then subjected to a carbodiimide coupling
reaction wherein VIIIa is treated with an acid R.sup.7' COOH in the
presence of WSC or DCC, and HOBT, and NMM, in the presence of an inert
organic solvent such as dimethylformamide (DMF), THF or
N-methylpyrrolidone, to form compound IXa. Alternatively, VIIIa may be
treated with R.sup.7' SO.sub.2 Cl and triethylamine or R.sup.7' OCOCl and
triethylamine to form Ixa.
Compound IXa is then treated with H.sub.2 /Pd--C or HBr/acetic acid (HOAc)
(where PG' is CBZ) to form compound of the invention Ia'. Ia' is separated
by conventional procedures and the desired isomers are guanylated with
amidine sulfonic acid, or other guanylation agent such as
1H-pyrazole-1-carboxamidine in the presence of an alcohol solvent such as
ethanol to form the compound of the invention Ib.
Referring to Reaction Sequence A, Part B1) compounds of formula Ia of the
invention wherein R.sup.6 is H and R.sub.b is --CO--A-- guanidine may be
prepared by deprotecting amide VIIa with TFA where PG' is BOC and PG is
not BOC, guanylating the deprotected amide by treatment with
amidinesulfonic acid (as described above with regard to Ia') and then
removing the remaining protecting group PG by treatment with H.sub.2
/Pd--C where PG is CBZ, to form compound Ic of the invention.
In Reaction Sequence A, Part B2) compounds of formula Ia of the invention
wherein R.sup.6 is H and R.sub.b is --CO--A--NH.sub.2 are prepared by
deprotecting Ia' with TFA (where R.sup.6 is BOC) or with H.sub.2 /Pd--C
(where R.sup.6 is CBZ) to form compound Id of the invention.
In Reaction Sequence A, Part C, compounds of formula Ia of the invention
where R.sub.b is --CO--A.sup.1 --R.sup.3a and R.sup.3a is amidine may be
prepared by treating amine IVa with acid IV" in the presence of WSC, HOBT
and triethylamine, at a temperature within the range of from about
-30.degree. to about 50.degree. C., to form nitrile Xa which is made to
undergo amidine formation by treating Xa with HCl in the presence of
methanol and then reacting the resulting compound with ammonia in the
presence of methanol to form amidine XIa. The protecting group, PG, is
removed in this process if PG=BOC. Amidine XIa is then made to undergo a
carbodiimide coupling reaction with protected amino acid VI" in the
presence of WSC or DCC, and HOBT and NMM (as described above with respect
to amide IVa) to form amidine XIIa which is deprotected with TFA (where
PG' is BOC) to form compound Ie of the invention where R.sup.6 is H.
Amidine Ie may be coupled with coupling agent R.sup.7' COOH, R.sup.7'
OCOCl, or R.sup.7' SO.sub.2 Cl (as described above with respect to amide
VIIIa) to form amidine compound If of the invention where R.sup.6 is not
H.
In Reaction Sequence B, Part A compounds of formula Ia of the invention
where R.sub.b is --A'R.sup.3a and R.sup.3a is amino or guanidine are
prepared starting with amine IVa which is acylated by treating amine IVa
with trifluoroacetic anhydride in the presence of an inert organic solvent
such as dichloromethane, chloroform or ether and a weak organic base such
as pyridine, triethylamine or NMM, to form trifluoroacetamide XIIIa which
is alkylated by treating XIIIa with bromoalkylphthalimide XIII' in the
presence of a base such as an alkali metal carbonate, and an inert organic
solvent DMSO or DMF, to form protected phthalimide derivative XIVa. The
phthalimide derivative XIVa is deprotected, for example, by treatment with
TFA where PG is BOC, to form phthalimide derivative XVa which is coupled
with amino acid derivative XV' in the presence of WSC or DCC, and HOBT and
NMM as described hereinbefore in Reaction Sequence A with respect to amine
IVa, to form phthalimide derivative XVIa. Phthalimide derivative XVIa is
then deprotected by treatment with hydrazine or methyl hydrazine in the
presence of an alcohol solvent such as ethanol, and then with base such as
potassium carbonate or cesium carbonate, and methanol to form amine
compound Ig of the invention. Ig may then be guanylated as described
hereinbefore in Sequence A, Part A with respect to Ia', to form guanidine
compound Ih of the invention.
Compounds of formula Ia of the invention wherein R.sup.6 is H and R.sub.b
is --A.sup.1 --R.sup.3a and R.sup.3a is guanidine or amine are prepared as
outlined in Reaction Sequence B, Part B1) and 2), respectively, by
deprotecting Ih (where R.sup.6 is CBZ) or deprotecting Ig (where R.sup.6
is CBZ), with H.sub.2 /Pd--C to form guanidine compound Ij and amino
compound Ik of the invention.
In Reaction Sequence B, Part C, compounds of formula Ia where R.sub.b is
--A'R.sup.3a and R.sup.3a is amidine are prepared by alkylating compound
XIIIa with nitrile XIII" in the presence of a base such as an alkali metal
carbonate like cesium carbonate, to form nitrile XVIIa which is
deprotected with TFA (where PG is BOC) and then subjected to a
carbodiimide coupling reaction with amino acid XV' and WSC or DCC, and
HOBT and NMM as described hereinbefore with respect to VIa in Sequence A,
Part A, to form nitrile XVIIIa. Nitrile XVIIIa is then treated with HCl
and ammonia in the presence of an alcohol solvent such as ethanol to form
amidine compound I1 of the invention. If in compound XVIIIa, R.sup.6 is
BOC, then in the final product I1, R.sup.6 will be H.
Compounds of formula Ia of the invention wherein R.sub.b is --SO.sub.2
--A.sup.1 --R.sup.3a and R.sup.3a is guanidine are prepared as outlined in
Reaction Sequence C, Part A starting with azide IIIa which is deprotected
with TFA (where PG is BOC) and then subjected to a carbodiimide coupling
reaction by treating the so-formed, deprotected compound with amino acid
XV' in the presence of WSC or DCC, and HOBT and NMM (as described in
Sequence A for VIa) to form azide XIXa. Azide XIXa is reduced by treating
XIXa with H.sub.2 /Pd--C or other reducing agent as described in Sequence
A with respect to the reduction of IIIa) to form the corresponding amine
which is sulfonylated by treatment with the sulfonyl chloride XIX' in the
presence of dichloromethane or other inert solvent such as chloroform or
ether, and a weak organic base such as triethylamine or pyridine, at a
temperature of within the range of from about -30.degree. to about
50.degree. C., to form bromosulfonamide XXa. Compound XXa is then reacted
with sodium azide in the presence of an inert organic solvent such as DMF
or DMSO to form azide XXIa which is deprotected and then guanylated (as
described with respect to IXa in Sequence A, Part A) to form guanidine
compound Im of the invention. Im may then be deprotected where R.sup.6 is
CBZ or BOC as shown to form compound In of the invention.
In Reaction Sequence C, Part B1) the preparation of compounds of formula I
wherein R.sub.b is --SO.sub.2 --A.sup.1 --NH.sub.2 and R.sup.6 is H is
shown starting with azide XXIa which is reduced to form the amine Io where
R.sup.6 is H.
In Part B2), compounds of formula Ia wherein R.sub.b is --SO.sub.2
--A.sup.1 --NH.sub.2 and R.sup.6 is other than H is formed starting with
azide XXIa which is deprotected and then coupled with R.sup.7' COOH,
R.sup.7' SO.sub.2 Cl or R.sup.7' OCOCl (as described above in Sequence A
with respect to VIlla) to form amine Ip.
Compounds of formula Ia of the invention wherein R.sub.b is --SO.sub.2
--A.sub.1 -amidine are prepared as shown in Reaction Sequence C Part C by
subjecting compound XXa to a displacement reaction by reacting XXa with
cyanide to form nitrile XXIIa which is then amidinated by treatment with
HCl and NH.sub.3 in the presence of ethanol (as described hereinbefore in
Sequence A, Part C with respect to IVa) to form amidine Iq of the
invention wherein if R.sup.6 in XXIIIa is BOC, then R.sup.6 in Iq is H.
Preparation of compounds of formula Ia of the invention wherein R.sub.b is
--A.sup.3 --R.sup.4 and A.sup.4 is amidine is outlined in Reaction
Sequence D, Part A starting with alcohol XXIVa which is oxidized by
treatment with, for example, oxalyl chloride or other oxidizing agent such
as pyridine.SO.sub.3 or pyridinum dichromate, in the presence of an inert
organic solvent such as DMSO, ether or methylene chloride, and a weak
organic base such as triethylamine to form aldehyde XXVa which is made to
undergo reductive amination by treating xxVa with NaCNBH.sub.3 in the
presence of methanol, or NaB(OAc).sub.3 H in the presence of acetic acid
and dichloroethylene, and nitrile XXV' to form nitrile XXVIa. Nitrile
XXVIa is reacted with HCl and then ammonia in the presence of ethanol to
form amidine XXVIIa wherein the protecting group PG is removed if PG=BOC.
So-formed amidine XXVIIa is made to undergo a carbodiimide coupling
reaction by treating XXVIIa with amino acid coupling agent VI" in the
presence of WSC or DCC, and HOBT (as described hereinbefore in Sequence A,
Part A with respect to VIa) to form amidine XXVIIIa which is deprotected
to form amidine compound Ir of the invention where R.sup.6 is H.
Amidine Ir may be subjected to a coupling reaction with R.sup.7' COOH,
R.sup.7' SO.sub.2 Cl or R.sup.7' OCOCl (as described hereinbefore in
Sequence A, Part A with respect to VIIIa) to form amidine compound Is of
the invention wherein R.sup.6 is other than H.
Compounds of formula Ia of the invention wherein R.sub.b is --A.sup.3
--CH.sub.2 NH.sub.2 are prepared as shown in Reaction Sequence D, Part B
wherein aldehyde XXVa is subjected to a reductive amination employing
protected amine XXV' and the procedure as described in Sequence D, Part A
with respect to aldehyde XXVa to form protected amine XXIXa which is
deprotected and coupled with coupling agent VI' (as described in Sequence
A, Part A with respect to VIa) to form protected amine XXXa which is
deprotected to form compound XXXIa. Compound XXXIa may then be deprotected
to form amine compound It of the invention where R.sup.6 is H or XXXIa may
be made to undergo a coupling reaction (as described in Sequence A, Part A
with respect to VIIIa) to form amine compound Iu of the invention.
In Reaction Sequence D, Part C compounds of formula I wherein R.sub.b is
--A.sup.3 --R.sup.4 and R.sup.4 is guanidine are prepared starting with
XXVa which is deprotected and subjected to a carbodiimide coupling with
amino acid XV' (as described hereinbefore in Sequence B, Part C) to form
aldehyde XXXIIa. Aldehyde XXXIIa is then made to undergo reductive
amination with amine XXXII' (using procedures as described in Sequence D,
Part A with respect to XXIa) to form compound XXXIIIa which is deprotected
and guanylated (as described in Sequence A, Part A with respect to Ia') to
form guanidine compound Iw of the invention. Where R.sup.6 in Iw is CBZ,
Iw may be reduced with H.sub.2 /Pd--C to form guanidine compound Iy of the
invention.
As seen in Reaction Sequence E1, compounds of formula Iz and Iz' may be
prepared by deprotecting previously described alcohol XXIVa and coupling
the free amine with a protected amino acid to give compound XXXIVa. The
alcohol XXXIVa is oxidized to aldehyde XXXVa as described previously in
the preparation of compound XXVa. The aldehyde is coupled with a compound
of formula XXXVIa using the reductive amination methodology described in
the preparation of compound XXIXa to give compound Iz. In the case where
R.sup.6 =H, Iz can be deprotected (Pd--C/H.sub.2 if R.sup.6 =CBZ) to
provide compound Iz'. Compounds of formula Iz.sup.2 and Iz.sup.3 can be
prepared by reduction of azide XIXa as described in Reaction Sequence C
Part A to give the amine XXXVIIa. This amine is coupled to an acid of
formula XXXVIIIa using standard carbodiimide coupling methodology
described in the preparation of Va to give a compound of formula XXXIXa.
Compound XXXIXa is deprotected (TFA if PG=BOC) and guanylated with a
reagent such as 1H-pryrazole-1-carboxamidine to provide a compound of
formula Iz.sup.2. In the case where R.sup.6 =CBZ, the compounds of formula
Iz.sup.2 can be deprotected to give a compound of formula Iz.sup.3 where
R.sup.6 =H.
DESCRIPTION OF THE FOURTH EMBODIMENT OF THE INVENTION
In a fourth aspect or embodiment, the present invention relates to
heterocyclic thrombin inhibitors of the invention have the structure A. A.
##STR54##
including all stereoisomers thereof, and including all pharmaceutically
acceptable salts thereof; wherein n is 0, 1 or 2;
p is 0, 1 or 2;
##STR55##
A is aryl or cycloalkyl, or an azacycloalkyl ring A of 4 to 8 members in
the ring or an azaheteroalkyl ring A of 4 to 8 members in the ring, as
given by the structure
##STR56##
X is CH.sub.2, O, S or NH; q=0, 1, 2, 3 or 4 if X=CH.sub.2 ;
q=2, 3 or 4 if X=O, S, NH; and
Y.sup.1 and Y.sup.2 are independently H, alkyl or halo;
R.sup.4 is guanidine, amidine or aminoethyl;
R, R.sup.1 and R.sup.2 are as defined hereinbefore; and
R.sub.3 is hydrogen,
##STR57##
or --CO.sub.2 R.sup.7' (wherein R.sup.7' is lower alkyl, aryl, arylalkyl
or cycloheteroalkyl);
with the provisos that where A is aryl or cycloalkyl, R.sup.4 is guanidine,
amidine or aminomethyl;
where A is azacycloalkyl or azaheteroalkyl, R.sup.4 is amidine;
where X is a hetero atom (that is A is azaheteroalkyl), then there must be
at least a 2-carbon chain between X and any N atom in the ring A or
outside the ring A.
Examples of the A ring (azacycloalkyl or azaheteroalkyl) which may be
employed herein include
##STR58##
and the like.
Preferred are compounds of formula I wherein
n is 0 or 1,
R.sub.3 is H; R is aralkyl or hydroxyalkyl,
R.sup.1 and R.sup.2 are each H, p is 0 or 1,
Q is a single bond, A is an azacycloalkyl ring
##STR59##
where q is 1 or 2; R.sup.4 is amidine.
Most preferred are compounds of formula I wherein R.sub.3 is H, n is 0,
R.sup.1 and R.sup.2 are each H,
R is aralkyl such as benzyl,
p is 1, Q is a single bond, AR.sup.4 is
##STR60##
The compounds of formula A of the invention (fourth embodiment) may be
prepared according to the following reaction sequences.
The compounds of formula A of the invention wherein A is azacycloalkyl or
azaheteroalkyl and R.sup.5 is amidine may be prepared according to the
following Reaction Sequence a.
The compounds of formula B or C are known in the art or may be prepared by
those skilled in the art employing conventional preparatory techniques.
##STR61##
As seen in the above Reaction Sequence a., compounds of formula I wherein A
is azacycloalkyl or azaheteroalkyl, are prepared as follows. The protected
acid B is made to undergo a carbodiimide coupling reaction with amine C in
the presence of ethyl 3-(3-dimethylamino)propyl carbodiimide hydrochloride
(WSC) or dicyclohexylcarbodiimide (DCC), and 1-hydroxybenzotriazole
monohydrate (HOBT), and N-methylmorpholine (NMM), and in the presence of
an inert organic solvent such as dimethylformamide (DMF), THF or
N-methylpyrrolidone, to form the amide D. Amide D is deprotected by
treatment with, for example, H.sub.2 /Pd-C if p.sup.1 is CBz, to form
amine E. Amine E is treated with amidinesulfonic acid in the presence of
alcohol solvent, such as ethanol to give cyclic guanidine AI. Guanidine AI
is deprotected by treatment with trifluoroacetic acid (if R.sup.3 =BOC)
with or without the presence of dry inert organic solvent such as
dichloromethane, chloroform or THF at temperatures within the range of
from about -15.degree. to about 20.degree. C. to form amidine compound of
the invention BI.
The compounds of the invention where A is aryl or cycloalkyl and R.sup.4 is
amidine or guanidine may be prepared according to the following Reaction
Sequence b.:
##STR62##
As seen in Reaction Sequence b., compounds of formula A. where A is aryl or
cycloalkyl and R.sup.4 is amidine or guanidine are prepared as follows.
The protected acid B is subjected to a carbodiimide coupling reaction
wherein B is treated with protected amine CA in the presence of WSC or
DCC, and HOBT, and NMM, in the presence of an inert organic solvent such
as dimethylformamide, THF or N-methylpyrrolidone, to form amide CI. The
amide CI is then dissolved in an alcohol solvent such as ethanol or
methanol, to which HCl has been added and the mixture is hydrogenated over
Pd--C or Pd(OH).sub.2 --C in the case where R.sub.3 is carbobenzyloxy, to
form compound DI of the invention.
The compounds of formula A. of the invention wherein A is aryl or
cycloalkyl and R.sup.4 is aminomethyl may be prepared according to the
following Reaction Sequence c.:
##STR63##
As seen in the above Reaction Sequence c. compounds of formula A wherein A
is aryl or cycloalkyl and R.sup.4 is aminomethyl are prepared as follows.
The protected acid B is made to undergo a carbodiimide coupling reaction
with protected amino acid CB in the presence of ethyl 3-(3-dimethylamino
propyl carbodiimide hydrochloride (WSC) or dicyclohexylcarbodiimide (DCC),
and 1-hydroxybenzotriazole monohydrate (HOBT), and N-methylmorpholine
(NMM), and in the presence of an inert organic solvent such as
dimethylformamide (DMF), THF or N-methylpyrrolidone, to form the amide DA.
Amide DA is deprotected by treatment with trifluoroacetic acid (TFA) when
p.sup.1 is t-butoxycarbonyl (BOC) or H.sub.2 --Pd/C when p.sup.1 is
carbobenzyloxy (CBz), with or without the presence of dry inert organic
solvent such as dichloromethane, chloroform or THF, at temperatures within
the range of from about -15.degree. to about 20.degree. C. to form amide
EI of the invention.
The starting acid B may be prepared according to the following reaction
sequence:
##STR64##
As seen in the above reaction sequence, compounds of formula B are prepared
as follows. The ester L is made to undergo a carbodiimide coupling
reaction with protected amino acid E in the presence of ethyl
3-(3-dimethylamino)propyl carbodiimide hydrochloride (WSC) or
dicyclohexylcarbodiimide (DCC), and 1-hydroxybenzotriazole monohydrate
(HOBT), and N-methylmorpholine (NMM), and in the presence of an inert
organic solvent such as dimethylformamide (DMF), THF or
N-methylpyrrolidone, to form the amide M. Amide M is hydrolyzed by
treatment with base such as NaOH, KOH or LiOH to form an alkali metal salt
which is neutralized with strong acid such as HCl or oxalic acid to form
B.
DESCRIPTION OF THE FIFTH EMBODIMENT OF THE INVENTION
In a fifth aspect or embodiment, the present invention relates to
alkylsulfonamido heterocyclic thrombin inhibitors invention having the
structure (i)
##STR65##
wherein Gx is an amido moiety which is
##STR66##
including all stereoisomers thereof; and including all pharmaceutically
acceptable salts thereof; wherein Alkyl.sub.1-2 is methyl or ethyl and
R, R.sup.1, R.sup.2, n, m and Y are as defined hereinbefore with respect to
the formula I compound (first embodiment of the invention).
Preferred are compounds of formula (i) wherein n is 0 or 1; m is 2;
Alkyl.sub.1-2 is methyl; R is arylalkyl; R.sup.1 and R.sup.2 are
independently hydrogen or lower alkyl such as methyl or ethyl; and Y is
--NH--.
The compounds of formula (i) of the invention wherein Y is NH may be
prepared according to the following Reaction Sequence (i).
##STR67##
The compounds of formula I of the invention wherein Y is NH may also be
prepared according to the following Reaction Sequence (ii)
##STR68##
As seen in the above Reaction Sequence (i), compounds of formula I(i)
wherein Y is --NH--, are prepared as follows. The ester II(i) is made to
undergo a carbodiimide coupling reaction with protected amino acid III(i)
in the presence of ethyl 3-(3-dimethyl-amino)propyl carbodiimide
hydrochloride (WSC) or dicyclohexylcarbodiimide (DCC), and
1-hydroxybenzotriazole monohydrate (HOBT), and N-methylmorpholine (NMM),
and in the presence of an inert organic solvent such as dimethylformamide
(DMF), THF or N-methylpyrrolidone, to form the amide IV(i). Amide IV(i) is
deprotected by treatment with trifluoroacetic acid with or without the
presence of dry inert organic solvent such as dichloromethane, chloroform
or THF at temperatures within the range of from about -15.degree. to about
20.degree. C. Sulfonyl chloride V(i) is added followed by organic base
such as triethylamine, pyridine or N,N-diisopropylethylamine to form the
sulfonamide VI(i). Sulfonamide VI(i) is hydrolyzed by treatment with
alkali metal base such as NaOH or LiOH in the presence of an alcohol
solvent such as methanol or ethanol. The reaction mixture is acidified
with HCl, KHSO.sub.4 or H.sub.2 SO.sub.4, to form acid VII(i). The acid
VII(i) is then subjected to a carbodiimide coupling reaction wherein
VII(i) is treated with protected amine VIII(i) in the presence of WSC or
DCC, and HOBT, and NMM, in the presence of an inert organic solvent such
as dimethylformamide, THF or N-methylpyrrolidone, to form sulfonamide
IX(i). The sulfonamide IX(i) is then dissolved in an alcohol solvent such
as ethanol or methanol, to which HCl has been added and the mixture is
hydrogenated over Pd--C or Pd(OH).sub.2 --C in the case where p.sup.1 is
carbobenzyloxy. The product is then treated with amidine sulfonic acid
X(i) in the presence of an alcohol solvent such as ethanol to form the
compound of the invention IA(i).
As seen in the above Reaction Sequence (ii), compounds of formula I(i)
wherein Y is --NH--, are also prepared as follows. The protected acid
IIA(i) is made to undergo a carbodiimide coupling reaction with protected
diamine VIII(i) in the presence of ethyl 3-(3-dimethylamino)propyl
carbodiimide hydrochloride (WSC) or dicyclohexylcarbodiimide (DCC), and
1-hydroxybenzotriazole monohydrate (HOBT), and N-methylmorpholine (NMM),
and in the presence of an inert organic solvent such as dimethylformamide
(DMF), THF or N-methylpyrrolidone, to form the amide IXA(i). Amide IXA(i)
is deprotected by treatment with trifluoroacetic acid (TFA) when P is
t-butoxycarbonyl (BOC) or H.sub.2 --Pd/C when P is carbobenzyloxy (CBz),
with or without the presence of dry inert organic solvent such as
dichloromethane, chloroform or THF, at temperatures within the range of
from about -15.degree. to about 20.degree. C. to form amide IXB(i). The
amide IXB(i) is then subjected to a carbodiimide coupling reaction wherein
IXB(i) is treated with protected amine III(i) in the presence of WSC or
DCC, and HOBT, and NMM, in the presence of an inert organic solvent such
as dimethylformamide, THF or N-methylpyrrolidone, to form amide IXC(i).
The amide IXC(i) is then dissolved in an alcohol solvent such as ethanol
or methanol, to which HCl has been added and the mixture is hydrogenated
over Pd--C or Pd(OH).sub.2 --C in the case where p.sup.1 is CBz or treated
with trifluoroacetic acid when p.sup.1 is BOC. The product is then treated
with amidine sulfonic acid X(i) in the presence of an alcohol solvent such
as ethanol to form IXD(i). Compound IXD(i) is deprotected by treatment
with TFA when P is BOC or by treatment with H.sub.2 --Pd/C when P is CBz,
as described above, and sulfonyl chloride V(i) is added followed by
organic base such as triethyl-amine, pyridine or N,N-diisopropylethylamine
to form the sulfonamide IA(i).
The compounds of formula I of the invention wherein Y is S may be prepared
according to the following Reaction Sequence (iii).
##STR69##
Referring to the above Reaction Sequence (iii), compounds of formula I(i)
wherein Y.dbd.S can be prepared as follows. The acid VII(i) is subjected
to a carbodiimide coupling reaction wherein VII(i) is treated with an
aminoalcohol XI(i) in the presence of WSC or DCC, HOBT, and NMM, in the
presence of an inert organic solvent such as dimethylformamide, THF or
N-methylpyrroli-done, to form sulfonamide alcohol XII(i). The sulfonamide
alcohol XII(i) is reacted with p-toluene-sulfonyl chloride (TsCl) in
pyridine, or in a solvent such as methylene chloride or chloroform, with
N,N-dimethylaminopyridine to provide toluenesulfonate XIII(i). The
compound IB(i) (Y=S) is prepared by treating XIII(i) with thiourea in a
solvent such as DMF or DMSO at temperatures within the range of from about
25.degree. C. to about 100.degree. C.
IN GENERAL
The term "lower alkyl" or "alkyl" as employed herein by itself or as part
of another group includes both straight and branched chain radicals of up
to 18 carbons, preferably 1 to 8 carbons, such as methyl, ethyl, propyl,
isopropyl, butyl, t-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl,
4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl,
dodecyl, the various branched chain isomers thereof, and the like as well
as such groups including 1, 2 or 3 halo substituents (for example, to form
CF.sub.3 or CF.sub.3 CH.sub.2) and/or 1 or 2 of the following
substituents: an aryl substituent (for example, to form benzyl or
phenethyl), an alkyl-aryl substituent, a haloaryl substituent, a
cycloalkyl substituent, an alkylcycloalkyl substituent, an alkenyl
substituent, an alkynyl substituent, hydroxy or a carboxy substituent. It
will be appreciated that the same "alkyl" group may be substituted with
one or more of any of the above substituents.
The term "cycloalkyl" by itself or as part of another group includes
saturated cyclic hydrocarbon groups containing 3 to 12 carbons, preferably
3 to 8 carbons, which include cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl and cyclododecyl, any of
which groups may be substituted with substituents such as halogen, lower
alkyl, alkoxy, and/or hydroxy groups.
The term "aryl" or "Ar" as employed herein by itself or as part of another
group refers to monocyclic or bicyclic aromatic groups containing from 6
to 10 carbons in the ring portion, such as phenyl, or naphthyl. Aryl (or
Ar), phenyl or naphthyl may include substituted aryl, substituted phenyl
or substituted naphthyl, which may include 1 or 2 substituents on either
the Ar, phenyl or naphthyl such as lower alkyl, cyano, amino, alkylamino,
dialkylamino, nitro, carboxy, carboalkoxy, trifluoromethyl, halogen (Cl,
Br, I or F), lower alkoxy, arylalkoxy, hydroxy, alkylthio, alkylsulfinyl,
alkylsulfonyl, arylthio, arylsulfinyl and/or arylsulfonyl.
In a separate embodiment of the invention, where R.sup.3 in formula I is
phenyl, the phenyl group may include 3, 4 or 5 substituents such as alkyl,
for example, pentamethyl and 2,4,6-tri-isopropyl, and halo, for example,
pentafluoro.
The term "aralkyl", "aryl-alkyl" or "aryl-lower alkyl" as used herein by
itself or as part of another group refers to lower alkyl groups as
discussed above having an aryl substituent, such as benzyl.
The term "lower alkoxy", "alkoxy" or aralkoxy"includes any of the above
lower alkyl, alkyl or aralkyl groups linked to an oxygen atom.
The term "halogen" or "halo" as used herein by itself or as part of another
group refers to chlorine, bromine, fluorine or iodine with chlorine being
preferred.
The term "lower alkenyl" or "alkenyl" as employed herein by itself or as
part of another group includes a carbon chain of up to 16 carbons,
preferably 3 to 10 carbons, containing one double bond which will be
separated from "N" by at least one saturated carbon moiety such as
--(CH.sub.2).sub.q -- where q can be 1 to 14, such as 2-propenyl,
2-butenyl, 3-butenyl, 2-pentenyl, 4-pentenyl and the like, and may include
a halogen substituent such as I, Cl, or F.
The term "lower alkynyl" or "alkynyl" as employed herein by itself or as
part of another group includes a carbon chain of up to 16 carbons,
preferably 3 to 10 carbons, containing one triple bond which will be
separated from "N" by at least one saturated carbon moiety such as
--(CH.sub.2).sub.q' -- where q' can be 1 to 14, such as 2-propynyl,
2-butynyl, 3-butynyl and the like.
The term "heteroaryl" or heteroaromatic by itself or as part of another
group refers to a 5- to 10-membered aromatic ring(s) which includes 1, 2,
3 or 4 hetero atoms such as nitrogen, oxygen or sulfur, such as
##STR70##
and the like. The heteroaryl rings may optionally be fused to aryl rings
defined previously. The heteroaryl rings may optionally include 1 or 2
substituents such as halogen (Cl, Br, F or CF.sub.3), lower alkyl, lower
alkoxy, carboxy, amino, lower alkylamino and/or dilower alkylamino.
The term "cycloheteroalkyl" as used herein refers to a 5-, 6- or 7-membered
saturated ring which includes 1 or 2 hetero atoms such as nitrogen, oxygen
and/or sulfur, such as
##STR71##
and the like.
The term "azaheteroalkenyl" as used herein refers to a 4- to 8-membered
ring which includes 1 or 2 hetero atoms such as nitrogen, oxygen and/or
sulfur, such as
##STR72##
The term "amino acid side chain" refers to any of the known alpha-amino
acids such as arginine, histidine, alanine, glycine, lysine, glutamine,
leucine, valine, serine, homoserine, allothreonine, naphthylalanine,
isoleucine, phenylalanine and the like.
The compounds of formulae I, l., Ia, A and (i) of the invention can be
obtained as pharmaceutically acceptable acid addition salts by reacting a
free base with an acid, such as hydrochloric, hydrobromic, hydroiodic,
nitric, sulfuric, phosphoric, acetic, citric, maleic, succinic, lactic,
tartaric, gluconic, benzoic, methanesulfonic, ethanesulfonic,
benzenesulfonic, p-toluenesulfonic acid or the like.
The compounds of the present invention are serine protease inhibitors, and
in particular may inhibit thrombin, Factor Xa, and/or trypsin. The
compounds of the present invention are useful for the treatment or
prophylaxis of those processes which involve the production and/or action
of thrombin. This includes a number of thrombotic and prothrombotic states
in which the coagulation cascade is activated which include, but are not
limited to, deep vein thrombosis (DVT), disseminated intravascular
coagulopathy (DIC), Kasabach-Merritt syndrome, pulmonary embolism,
myocardial infarction, stroke, thromboembolic complications of surgery
(such as hip replacement and endarterectomy) and peripheral arterial
occlusion. In addition to its effects on the coagulation process, thrombin
has been shown to activate a large number of cells (such as neutrophils,
fibroblasts, endothelial cells, smooth muscle cells). Therefore, the
compounds of the present invention may also be useful for the treatment or
prophylaxis of adult respiratory distress syndrome, septic shock,
septicemia, inflammatory responses which include, but are not limited to,
edema, acute or chronic atherosclerosis, and reperfusion damage.
The compounds of the invention may also be useful in treating
neoplasia/metastasis (in particular those which utilize fibrin) and
neurodegenerative diseases such as Alzheimer's disease and Parkinson's
disease. In addition, the compounds of the present invention may be useful
to prevent restenosis following arterial injury induced by endogenous
(rupture of an atherosclerotic plaque) or exogenous (invasive
cardiological procedure) events.
The compounds of the present invention may also be used as an anticoagulant
in extracorpeal blood circuits, such as those necessary in dialysis and
surgery (such as coronary artery bypass surgery).
The compounds of the present invention may also be used in combination with
thrombolytic agents, such as tissue plasminogen activator (natural or
recombinant), streptokinse, urokinase, prourokinase, anisolated
streptokinase plasminogen activator complex (ASPAC), animal salivary gland
plasminogen activators, and the like. The compounds of the present
invention may act in a synergistic fashion to prevent reocclusion
following a successful thrombolytic therapy and/or reduce the time to
reperfusion. The compounds of the present invention may also allow for
reduced doses of the thrombolytic agent to be used and therefore minimize
potential hemorrhagic side-effects.
The compounds of the present invention may also be used in combination with
other antithrombotic or anticoagulant drugs such as thromboxane receptor
antagonists, prostacyclin mimetics, phosphodiesterase inhibitors,
fibrinogen antagonists, and the like.
Compounds of the present invention that inhibit trypsin may also be useful
for the treatment of pancreatitis.
The compounds of the invention can be administered orally or parenterally
to various mammalian species known to be subject to such maladies, e.g.,
humans, cats, dogs and the like in an effective amount within the dosage
range of about 0.1 to about 100 mg/kg, preferably about 0.2 to about 50
mg/kg and more preferably about 0.5 to about 25 mg/kg (or from about 1 to
about 2500 mg, preferably from about 5 to about 2000 mg) on a regimen in
single or 2 to 4 divided daily doses.
The active substance can be utilized in a composition such as tablet,
capsule, solution or suspension containing about 5 to about 500 mg per
unit of dosage of a compound or mixture of compounds of formula I, l., Ia
or A. They may be compounded in conventional matter with a physiologically
acceptable vehicle or carrier, excipient, binder, preservative,
stabilizer, flavor, etc., as called for by accepted pharmaceutical
practice.
The following Examples represent preferred embodiments of the first
embodiment of the present invention. Unless otherwise indicated, all
temperatures are expressed in degrees Centigrade.
EXAMPLE 1
1(S),2.alpha.,4.beta.!-N-4-(Aminoiminomethyl)
amino!butyl!-1-3-hydroxy-2-(2-naphthalenylsulfonyl)amino!-1-oxopropyl!-4
-methyl-2-piperidinecarboxamide, trifluoroacetate (1:1) salt
A. (2R-trans)-1-(Phenylmethoxy)carbonyl!-4-methyl-2-piperidinecarboxylic
acid, ethyl ester
A(1). 4-Methyl-2-piperidinecarbonitrile
To 500 g of sodium hypochlorite solution (5% in Cl, Aldrich) cooled in an
ice bath was added dropwise 33.6 g (340 mmol, Aldrich) of
4-methylpiperidine, over 40 minutes. The reaction mixture was stirred for
20 minutes then poured into a separatory funnel and extracted with two-400
mL portions of ether. The extracts were combined, dried (sodium sulfate)
and concentrated invacuo to give .about.44 g of N-chloro intermediate as a
yellow liquid.
To a solution of 19.1 g (340 mmol) of potassium hydroxide in 140 mL of 95%
ethanol heated to 80.degree. was added dropwise over 40 minutes the
solution of crude N-chloropiperidine from above in 20 mL of 95% ethanol.
The addition was mildly exothermic and a precipitate formed. The reaction
mixture was stirred for 10 minutes, cooled to room temperature,
concentrated in vacuo, and to the residue was added 85 mL of 2N aqueous
NaOH solution. The resulting mixture was extracted with three 75 mL
portions of ether. The ether extracts were combined, dried (sodium
sulfate) and concentrated in vacuo to give the crude imine as a viscous
yellow oil.
To a solution of 110 g (1.7 mol, Mallinckrodt) of potassium cyanide in 400
mL of water cooled in an ice-bath was added over .about.1 h 183 mL (2.20
mol) of concentrated HCl, followed by the crude imine from above. The
reaction mixture was stirred between 10.degree.-20.degree. C. for 4 h then
cooled in an ice-bath and basified to pH 12 by addition of .about.80 g of
potassium hydroxide pellets. the resulting solution was poured into a
separatory funnel and extracted with three-300 mL portions of ether. The
ether layers were combined, dried (sodium sulfate) and concentrated
invacuo to give a yellow oil. The crude material was purified by simple
distillation at reduced pressure to afford 13.2 g (106 mmol, 31%) of title
nitrile as a clear liquid, bp 72-74.degree. (3 mm).
A(2). 4-Methyl-2-piperidinecarboxylic acid, ethyl ester
To 250 mL of 6N aqueous HCl solution was added 13.0 g (105 mmol) of Part
A(1) nitrile at room temperature. The reaction mixture was heated to
reflux (bath temp 145.degree.) for 6 h then cooled to room temperature and
concentrated invacuo to give the crude acid hydrochloride as a solid.
To 250 mL of absolute ethanol cooled in an ice-bath was added dropwise 40
mL (550 mmol) of thionyl chloride over 30 minutes. The solution was
stirred for an additional 15 minutes then added to the crude acid
hydrochloride from above. The resulting slurry was heated to reflux for 4
h then cooled to room temperature and filtered to remove solids. The
filtrate was concentrated in vacuo to give the crude ester amine
hydrochloride as brown oil. The oil was partitioned between 150 mL of
saturated aqueous potassium carbonate solution and 150 mL of chloroform.
The aqueous layer was separated and extracted with two-100 mL portions of
chloroform. The organic extracts were combined, dried (sodium sulfate) and
concentrated to give crude title ester as a brown oil. The crude material
was purified by distillation through a 10 cm packed column (glass helices,
3 mm) at reduced pressure to afford 11.0 g (64.3 mmol, 61%) of title amine
as a colorless liquid, bp 37.degree.-38.degree. (0.4 mm). The trans/cis
ratio was determined as .about.6:1 by 270 MHz .sup.1 H NMR.
A(3).
(2R-trans)-1-(Phenylmethoxy)carbonyl!-4-methyl-2-piperidinecarboxylic aci
d, ethyl ester
To a solution of 10.0 g (58.5 mmol) of Part A(2) amine in 100 mL of
methylene chloride cooled to 0.degree. was added 9.8 mL (70 mmol,
distilled from calcium hydride) of triethylamine in one portion then
dropwise 11.9 g (70 mmol, Aldrich) of benzyl chloroformate over 20
minutes. The reaction mixture was stirred for 30 minutes then washed with
100 mL of 1N aqueous HCl, 100 mL of saturated aqueous sodium bicarbonate
solution, 50 mL of brine, dried (magnesium sulfate) and then concentrated
in vacuo to give an oil. The crude oil was purified by flash
chromatography (Merck silica, 30.times.10 cm, 600 g, 1:9 EtOAc/hexane) to
afford 12.7 g (41.6 mmol, 71%) of title compound (trans isomer) as a
colorless oil. In addition 2.95 g (9.67 mmol, 17%) of a .about.1:1 mixture
of trans/cis isomers was obtained as a colorless oil.
B. (2R-trans)-4-Methylpiperidinecarboxylic acid, ethyl ester
Part A racemic CBZ ester (2.36 g, 7.73 mmol) was dissolved in absolute
ethanol (50 mL) and hydrogenated over Pd--C (10%, 250 mg) at RT and 1 atm.
After 6 hrs additional catalyst (50 mg) was added, the reaction continued
for 2 hours, filtered, and evaporated in vacuo to provide the title free
amine (1.22 g, 7.13 mmol, 92%).
C.
(2R-trans)-1-N-(1,1-Dimethylethoxy)carbonyl!-O-(phenylmethyl)-L-seryl!-4
-methyl-2-piperidinecarboxylic, ethyl ester
Part B pipecolic ester (1.22 g, 7.13 mmol) and BOC-Ser(OBn)--OH (2.36 g,
8.0 mmol) were dissolved in DMF (15 mL) at RT. HOBT (1.08 g, 8.0 mmol),
WSC (1.54 g, 8.0 mmol) and N-methyl morpholine (NMM) (0.88 mL, 8.0 mmol)
were added. The pH was ca 8.5. The reaction was stirred for 90 min,
partitioned between ethyl acetate and 10% KHSO.sub.4, the aqueous layer
back-extracted, and the organic layers combined. The combined organic
layers were washed with saturated NaHCO.sub.3 (2.times.), saturated NaCl,
dried over magnesium sulfate and stripped in vacuo to provide title ester
as an oil (2.99 g, 6.67 mmol, 93%).
D.
(2R-trans)-1-N-(2-Naphthalenylsulfonyl)-O-(phenylmethyl)-L-seryl!-4-methy
l-2-piperidinecarboxylic acid, ethyl ester
Part C BOC-Ser-pipecolic ester (2.80 g, 6.25 mmol) was dissolved in
trifluoroacetic acid (TFA) at 0.degree. C., and allowed to warm to RT over
60 min. The TFA was evaporated in vacuo for two hours to provide the TFA
salt as an oil (4.2 g). The crude TFA salt was dissolved in
dichloromethane (15 mL) and 2-naphthylsulfonyl chloride (Aldrich, 1.58 g,
7.0 mmol) was added, followed by triethylamine (3.5 mL, 25 mmol). A
vigorous exotherm developed, and the reaction was put into an ice bath for
10 min, then warmed to RT. After 6 hr the dichloromethane was evaporated,
the reaction mixture partitioned between ethyl acetate and 10% KHSO.sub.4
(2.times.), washed with saturated NaHCO.sub.3 (2.times.), dried over
magnesium sulfate and evaporated to provide crude title ester (3.18 g).
The crude product was chromatographed on silica gel (1:3==>1:2 ethyl
acetate (EtOAc):hexanes to provide the two diastereomers as an oil (2.29
g, 4.25 mmol, 68% over two steps).
E.
(2R-trans)-1-N-(2-Naphthalenylsulfonyl)-O-(phenylmethyl)-L-seryl!-4-methy
l-2-piperidinecarboxylic acid
The Part D ester was dissolved in methanol (MeOH) (30 mL) and treated with
1N NaOH (10 mL, 10 mmol). The reaction was stirred for 12 hr, and
additional NaOH (5 mL) was added. After 24 hr the reaction was acidified
with HCl (50 mL, 1N), extracted with ethyl acetate (2.times.), and dried
in vacuo to give title free acid (1.99 g, 3.90 mmol, 99%).
F. (4-Aminobutyl)carbamic acid, phenylmethyl ester
1,4-butanediamine dihydrochloride (5.96 g, 37 mmol) was dissolved in water
(15 mL), and DMF (50 mL) was added. With rapid stirring, CBZ-Cl (1.0 mL,
7.0 mmol) was added dropwise over 2 minutes. The cloudy solution had pH
.about.2.8. The pH was adjusted to 9.0 with 5 E NaOH, (clear solution) and
stirred for 2 hr. The reaction mixture was acidified to pH=1.25, extracted
with ether (2.times.), made basic (pH>10) with 5 N NaOH, and extracted
with dichloromethane (2.times.). The dichloromethane fractions were
combined and washed with water (2.times.), dried and evaporated in vacuo
to provide the crude mono-CBZ amine (0.72 g, 3.24 mmol, 46%) which was
used as is in the next step.
G.
(2R-trans)-1-N-(2-Naphthalenylsulfonyl)-O-(phenylmethyl)-L-seryl!-N-4-
(phenylmethoxy)carbonyl!amino!butyl!-4-methyl-2-piperidinecarboxamide
To a solution of Part E mono-CBZ amine (400 mg, 1.8 mmol) and Part E acid
(640 mg, 1.25 mmol) in DMF (5 mL) was added HOBT (170 mg, 1.25 mmol), WSC
(0.25 g, 1.3 mmol), and NMM (138 .mu.L, 1.3 mmol). The pH was .about.8.5.
The reaction was stirred for two hr (complete by HPLC), partitioned
between ethyl acetate and 10% KHSO.sub.4, back-extracted, and the combined
organic layers washed with 10% KHSO.sub.4, saturated NaHCO.sub.3
(2.times.), saturated NaCl, and evaporated to provide the crude product in
quantitative yield. The crude product was chromatographed on silica gel
(2:1:EtOAc:hexanes) to provide a mixture of the title two diastereomers
(671 mg, 0.94 mmol, 75%).
H.
1(S),2.alpha.,4.beta.!-N-4-(Ainoiminomethyl)amino!butyl!-1-3-hydroxy-2
-(2-naphthalenylsulfonyl)amino!-1-oxopropyl!-4-methyl-2-piperidinecarboxam
ide, trifluoroacetate (1:1) salt
The Part G O-benzyl CBZ derivative was dissolved in ethanol (20 mL) to
which acetyl chloride had been added (0.71 mL, 10 mmol, gives a 0.5M
solution of HCl), and the mixture hydrogenated over Pd--C (10%, 150 mg) at
1 atm and reflux temperature. After 5 hr the reaction mixture was
filtered, stripped and fresh ethanol and catalyst added. The reaction was
determined to be ca 80% complete after an additional 2 hr, whereupon it
was filtered and stripped to provide the crude amine (412 mg). The crude
material was purified by preparative HPLC (80:20=>50:50) to provide the
des-benzyl, des-CBZ diastereomers (281 mg, 0.57 mmol, 71%) and the
O-benzyl, des-CBZ compounds (69 mg, 0.12 mmol, 15%).
The des-benzyl, des-CBZ material above (208 mg, 0.46 mmol) was dissolved in
ethanol (5 mL), to which amidinesulfonic acid (78 mg, 0.65 mmol) and
triethyl amine (176 .mu.L, 1.26 mmol) were added. After 20 hr the solvent
was evaporated and the material purified by preparative HPLC
(80:20=>50:50). Fractions containing the two major components were
combined and lyophilized to provide title compound as a 40:60 mixture of
isomers A:B (255 mg, 78%). Purity.gtoreq.98%.
.alpha.!.sub.D (c=1.0, MeOH)=-11.7.degree..
Analysis calc'd for 1.30 TFA+0.80 H.sub.2 O: C, 47.68; H, 5.64; N, 12.09;
F, 10.66. Found: C, 47.68; H, 5.34;, N, 11.97; F, 10.83.
EXAMPLE 2
1(S),2.alpha.,4.beta.!-N-4-(Aminoiminomethyl)amino!butyl!-4-methyl-1-2-
(2-naphthalenylsulfonyl)amino!-1-oxo-3-(phenylmethoxy)propyl!-2-piperidine
carboxamide, trifluoroacetate (1:1) salt
The O-benzyl, des-CBZ compound described in Example 1 Part H (69 mg, 0.099
mmol) was dissolved in ethanol (2 mL), to which amidinesulfonic acid (22
mg, 0.18 mmol, prepared as described in Synthesis (1986) 777-779), and
triethyl amine (50 .mu.L, 0.36 mmol) were added. After 20 hr the solvent
was evaporated and the material purified by preparative HPLC
(80:20=>50:50). Fractions containing the two major components were
combined and lyophilized to provide title compound as a 1:3 mixture of
isomers A:B (76 mg, 100%). Purity.gtoreq.98%.
.alpha.!.sub.D (c=1.0, MeOH)=-5.6.degree..
Analysis calc'd for 1.10 TFA+0.30 H.sub.2 O: C, 54.51; H, 5.84; N, 11.15;
F, 8.32. Found: C, 54.19; H, 5.84;, N, 11.04; F, 8.29.
EXAMPLE 3
2R-1(S*),2.alpha.,4.beta.!-N-4-(Aminoiminomethyl)amino!butyl!-1-3-hydr
oxy-2-(2-naphthalenylsulfonyl)amino!-1-oxopropyl!-4-methyl-2-piperidinecar
boxamide, trifluoroacetate (1:1) salt
Example 1 compound (ca 220 mg) was dissolved in water (15 mL) and
chromatographed on a YMC S-10 20 mm.times.500 mm ODS column, using an
80:20==>50:50 gradient. After lyophilization, the first peak to elute
(isomer A) comprised 95 mg. The stereochemistry (2R, 4R) was assigned by
comparison to a sample prepared form non-racemic starting material.
.alpha.!D (c=1.00, MeOH)+11.0.degree..
Analysis calc'd for 1.16 H.sub.2 O+1.07 TFA: C, 48.25; H, 5.88; N, 12.44;
F, 9.03; S, 4.75. Found: C, 48.25; H, 5.61; N, 12.25; F, 9.25; S, 5.19.
EXAMPLE 4
2S-1(R*),2.alpha.,4.beta.!-N-4-(Aminoiminomethyl)amino!butyl!-1-3-hydr
oxy-2-(2-naphthalenylsulfonyl)amino!-1-oxopropyl!-4-methyl-2-piperidinecar
boxamide, trifluoroacetate (1:1) salt
Example 1 compound (ca 220 mg) was dissolved in water (15 mL) and
chromatographed on a YMC S-10 20 mm.times.500 mm ODS column, using an
80:20==>50:50 gradient (acetonitrile:water). After lyophilization, the
second peak to elute (isomer B) comprised 103 mg. The stereochemistry (2S,
4S) was assigned by comparison of the two isomers.
.alpha.!D (c=1.00, MeOH)-29.0.degree..
Analysis calc'd for 1.39 H.sub.2 O+1.17 TFA: C, 47.52; H, 5.83; N, 12.16;
F, 9.65; S, 4.64. Found: C, 47.52; H, 5.49; N, 11.96; F, 9.60; S, 5.25.
High resolution MS shows (M+H).sup.+ =533.2546 (C.sub.25 H.sub.37 O.sub.5
N.sub.6 S).
EXAMPLE 5
(2S-trans)-N-4-(Aminoiminomethyl)amino!butyl!-4-methyl-1-(1,2,3,4-tetr
ahydro-3-methyl-8-quinolinyl)sulfonyl!amino!acetyl!-2-piperidinecarboxamide
, trifluoroacetate salt
A. (2S-trans)-4-Methyl-2-piperidinecarboxylic acid, ethyl ester
To a solution of dry EtOH (65 mL) and acetyl chloride (6.52 mL added
dropwise at 0.degree. C.) was added (2S, 4S)-4-methyl pipecolic acid (2.8
g, 19.6 mmol at room temperature. The reaction was stirred at room
temperature for 16 hrs and then concentrated it vacuo to give the title
compound, (3.01 g, 90%).
B.
(2S-trans)-1-(1,1-Dimethylethoxy)carbonyl!amino!acetyl!-4-methyl-2-pipe
ridinecarboxylic acid, ethyl ester
To a solution of L-Boc-Glycine (0.4 g, 2.25 mmol) and HOBT (0.31 g, 2.25
mmol) in 7 mL DMF was added WSC (0.44 g, 2.25 mmol) and Part A ethyl ester
(0.35 g, 2.04 mmol), followed by NMM (240 .mu.l, 2.25 mmol) to adjust the
pH of the solution to 8. The reaction was stirred at room temperature for
16 hrs and worked up by washing with a saturated solution of NaHCO.sub.3,
a saturated solution of KHSO.sub.4, brine, dried over Na.sub.2 SO.sub.4
and evaporated to give the title compound (0.60 g, 89%).
(M+H).sup.+ @329
C.
(2S-trans)-1-(1,1-Dimethylethoxy)carbonyl!amino!acetyl!-4-methyl-2-pipe
ridinecarboxylic acid
To a solution of Part B (560 mg, 1.71 mmol) in 6.5 mL EtOH was added 1N
NaOH (6.5 mL) at 0.degree. C. The reaction was stirred at room temperature
for 16 hrs and concentrated in vacuo. The reaction was acidified with 1N
HCl to pH 2 and extracted with EtOAc. Then the EtOAc was washed with a
saturated solution of KHSO.sub.4, brine, dried over Na.sub.2 SO.sub.4 and
concentrated to give the title compound (512 mg, quantitative).
(M+H).sup.+ @301
D. (4-Aminobutyl)carbamic acid, phenylmethyl ester
To a solution of 1,4-diaminopentane (25 g, 155 mmol) in 100 mL DMF/H.sub.2
O (1:1) was added 5N NaOH to adjust the pH to 9. CBZ-Cl (3.7 mL, 26 mmol)
was added as one portion and stirred at room temperature for 16 hrs. The
reaction was acidified to pH 1.25 and washed with diethyl ether. The pH of
the reaction was then adjusted to 9.5 and the reaction extracted with
EtOAc. The organic layer was washed with H.sub.2 O, brine, dried over
Na.sub.2 SO.sub.4 and evaporated to give title compound (1.57 g, 30%).
E.
(2S-trans)-1-(1,1-Dimethylethoxy)carbonyl!amino!acetyl!-4-methyl-N-4-
(phenylmethoxy)carbonyl!amino!butyl!-2-piperidinecarboxamide
To a solution of Part C compound (516 mg, 1.72 mmol) and HOBT (256 mg, 1.89
mmol) in 10 mL DMF was added WSC (363 mg, 1.89 mmol) and Part D CBZ amine,
(381 mg, 1.72 mmol), followed by NMM (198 .mu.l, 1.89 mmol) to adjust the
pH of the solution to 8. The reaction was stirred at room temperature for
16 hrs. The reaction was worked up by pouring into a saturated solution of
NaHCO3 and extracting with EtOAc. The EtOAc layer was washed with a
saturated solution of KHSO.sub.4, brine, dried over Na2SO.sub.4 and
concentrated in vacuo to give the title compound (900 mg, quantitative).
F.
(2S-trans)-1-(Aminoacetyl)-4-methyl-N-4-(phenylmethoxy)carbonyl!amino!b
utyl!-2-piperidinecarboxamide, hydrochloride
To a solution of Part E compound (0.76 g, 1.51 mmol) in 5 mL MeOH was added
4.5 mL of 4N HCl/dioxane at 0.degree. C. After 2 hrs at room temperature,
the reaction was worked up by adding Et.sub.2 O to precipitate the title
compound (0.70 g, quantitative).
G.
(2S-trans)-4-Methyl-1-(3-methyl-8-quinolinyl)sulfonyl!amino!acetyl!-N-
4-(phenylmethoxy)carbonyl!amino!butyl!-2-piperidinecarboxamide
To a solution of Part F compound (500 mg, 1.13 mmol) in 11 mL CHCl.sub.3
was added Et.sub.3 N (475 ml, 3.39 mmol) at 0.degree. C. After 5 minutes,
6-methyl-8-quinolinesulfonyl chloride, (273 mg, 1.13 mmol) was added. The
reaction was stirred at 0.degree. C. for another 5 minutes then stirred at
room temperature for 3 hrs. The reaction was washed with a saturated
solution of NaHCO.sub.3, a saturated solution of KHSO.sub.4, brine, dried
over Na.sub.2 SO.sub.4 and evaporated to give the crude product which was
purified by a silica gel column, using CH.sub.2 Cl.sub.2 :MeOH eluting
solvent to give the title compound (150 mg, 22%).
H.
(2S-trans)-N-(4-Aminobutyl)-4-methyl-1-(1,2,3,4-tetrahydro-3-methyl-8qu
inolinyl)sulfonyl!amino!acetyl!-2-piperidinecarboxamide, hydrochloride
A solution of Part G compound (140 mg, 0.23 mmol) and 230 .mu.l 1N HCl in 5
mL EtOH was hydrogenated at 1 atm over 10% Pd/C (40 mg). After 16 hrs, the
suspension was filtered through Celite and concentrated in vacuo to give
the title free amine, (130 mg, quantitative).
I.
(2S-trans)-N-4-(Aminoiminomethyl)amino!butyl!-4-methyl-1-(1,2,3,4-tet
rahydro-3-methyl-8-quinolinyl)sulfonyl!amino!acetyl!-2-piperidinecarboxamid
e
To a solution of Part H compound (80 mg, 0.145 mmol) in 0.8 mL EtOH was
added Et.sub.3 N (60 ml, 0.43 mmol), followed by amidinesulfonic acid (27
mg, 0.22 mmol). After 16 hrs at room temperature, the reaction was
concentrated in vacuo and was subjected to preparative HPLC using an 80:20
to 50:50 gradient (H.sub.2 O:CH.sub.3 CN) over 50 minutes to give the
title compound (40 mg, 50%).
(M+H)+@522
Anal Calc'd for C.sub.24 H.sub.39 N.sub.7 O.sub.4 S.1.2TFA.1.1 H.sub.2 O C,
43.54; H, 6.04; N, 13.46; F, 9.39 Found C, 43.87; H, 5.72; N, 13.09; F,
9.33.
.alpha.!.sub.D =-6.7 (C=0.51, methanol)
EXAMPLE f6
trans-N-4-(Aminoiminomethyl)amino!butyl!-4-(methylthio)-1-(2-naphthalen
ylsulfonyl)amino!acetyl!-L-prolinamide, trifluoroacetate salt
A. cis-4-Hydroxy-1-(phenylmethoxy)carbonyl!-L-proline, methyl ester
N-CBZ-L-keto proline was dissolved in methanol (150 mL), cooled to
0.degree. to 5.degree. C. and treated dropwise with a solution of
NaBH.sub.4 (2.875 g) in water (10 mL). The mixture was kept at 0.degree.
to 5.degree. C. for 19 h. The methanol was removed in vacuo and the
residue was treated with 75 mL of 3N NaOH and stirred at room temperature
for 40 min. After cooling in an ice bath, the mixture was acidified with
conc HCl. The product was extracted into ethyl acetate (3.times.50 mL).
The combined extracts were washed with brine (2.times.40 mL), dried
(MgSO.sub.4) and freed of solvent in vacuo to give the cis-hydroxy acid.
This was converted to the methyl ester by dissolving in methanol (75 mL),
treating with acetyl chloride (7.5 mL), and stirring overnight at room
temperature. The solvent was removed. The residue was dissolved in ethyl
acetate (75 mL), washed with sodium bicarbonate solution (2.times.25 mL),
dried (MgSO.sub.4) and freed of solvent in vacuo to give title compound as
a viscous oil (4.315 g, 81%).
B.
cis-4-(4-Methylphenyl)sulfonyl!oxy!-1-(phenylmethoxy)carbonyl!-L-prolin
e, methyl
The Part A alcohol (4.315 g, 16.84 mmol) was dissolved in dry pyridine (23
mL), cooled in an ice bath and treated with tosyl chloride (6.8 g). The
mixture was allowed to warm slowly to room temperature and left stirring
60 h. After cooling, cold 2N HCl was added and the mixture was left at
0.degree. to 5.degree. C. for 4 h. The solid was collected by filtration
and dissolved in dichloromethane. This solution was washed with 1N HCl
soln and brine, dried (MgSO.sub.4) and freed of solvent in vacuo leaving
title compound as a crystalline material.
C. 4-(Methylthio)-1-(phenylmethoxy)carbonyl!-L-proline, methyl ester
Part B tosylate (2.09 g, 4.8 mmol) was dissolved in absolute ethanol (20
mL) and acetone (20 mL), under an atmosphere of argon, and treated with
sodium thiomethoxide (1.39 g, 19.9 mmol). The mixture was heated under
reflux 1 h, cooled and most of the solvent was removed in vacuo. The
residue was partitioned between 1N HCl and ethyl acetate. The ethyl
acetate layer was washed with water, dried (MgSO.sub.4), and freed of
solvent in vacuo. NMR and TLC indicated the material was mainly the free
acid. It was converted to the methyl ester by dissolving in methanol (20
mL), adding acetyl chloride (2 mL) and stirring at room temperature for
two hours. After removal of the solvent, the product was purified by
chromatography on silica gel, eluting with ethyl acetate:hexane (1:2) to
give the title thiomethyl compound (1.364 g, 99%).
D. 4-(Methylthio)-L-proline, methyl ester, hydrobromide
Part C compound (945 mg, 3.3 mmol) was treated with 30% HBr in HOAc (8 mL)
for 1 h and then concentrated in vacuo. Trituration of the residue with
ether gave a solid which was harvested by filtration and washed with more
ether leaving the title deprotected amine (749 mg, 89%).
E.
trans-1-(1,1-Dimethylethoxy)carbonyl!amino!acetyl!-4-(methylthio)-L-pro
line, methyl and
F. cis-1-N-(1,1-Dimethylethoxy)carbonyl)glycyl!-4-(methylthio)-L-proline,
methyl ester
Part D L-proline derivative (1.012 g, 3.95 mmol) and BOC-glycine (900 mg,
5.1 mmol) were dissolved in DMF (20 mL) at RT. HOBT (688 mg, 5.1 mmol),
WSC (974 mg, 5.1 mmol) and NMM (1.0 mL, 9.1 mmol) were added. The reaction
was stirred for 16 h, partitioned between ethyl acetate(50 mL) and 10%
KHSO.sub.4 (100 mL). The aqueous layer was extracted with ethyl acetate
(2.times.50 mL), and the organic layers were combined. The combined
organic layers were washed with saturated NaHCO.sub.3, saturated NaCl,
dried over magnesium sulfate and concentrated in vacuo to provide an oil.
TLC indicated this was two major products. The crude material was purified
by chromatography on silica gel. Elution with ethyl acetate:hexanes (1:2
followed by 1:1) gave the major product (tentatively assigned the trans
configuration of the title compound, 603 mg, 46%) and the minor product
(tentatively assigned the cis configuration of the title compound, 204 mg,
16%) as well as some mixed fractions (438 mg, 33%).
G.
trans-4-(Methylthio)-1-(2-naphthalenylsulfonyl)amino!acetyl!-L-proline,
methyl ester
The Part E BOC derivative (600 mg, 1.8 mmol) was dissolved in
trifluoroacetic acid (TFA) (15 mL) and stirred at RT 1 h. The TFA was
removed by distillation under reduced pressure and by coevaporation with
toluene. The crude TFA salt was dissolved in dichloromethane (20 mL),
cooled in an ice bath, and 2-napthylsulfonyl chloride (Aldrich, 450 mg,
2.0 mmol) was added, followed by triethylamine (1.5 mL). The solution was
warmed to RT and stirred 1.5 h before diluting with dichloromethane (75
mL). This solution was washed with potassium hydrogen sulfate
solution(2.times.40 mL) and saturated NaHCO.sub.3 (2.times.40 mL), dried
over magnesium sulfate, and evaporated to provide crude title compound.
The crude product was chromatographed on silica gel and eluted with 50%
EtOAc in hexanes to provide title sulfonamide as a foam (700 mg, 92%
overall in two steps).
H. trans-4-(Methylthio)-1-(2-naphthalenylsulfonyl)amino!acetyl!-L-proline
A solution of Part G methyl ester (700 mg, 1.65 mmol) in methanol (20 mL)
was treated with 1N NaOH solution (8 mL) and stirred at room temperature
for three h. After acidification with 1N HCl solution, the product was
extracted into dichloromethane (2.times.50 mL), dried over magnesium
sulfate and freed of solvent in vacuo to give title compound as a white
foam (687 mg, 100%).
I.
trans-4-(Methylthio)-1-(2-naphthalenylsulfonyl)amino!acetyl!-N-4-(phe
nylmethoxy)carbonyl!amino!butyl!-L-prolinamide
To a solution of mono-CBZ butyl diamine (484 mg, 2.18 mmol) and Part H acid
(687 mg, 1.65 mmol) in DMF (25 mL) was added HOBT (230 mg, 1.7 mmol), WSC
(325 mg, 1.7 mmol), and NMM (373 .mu.L, 3.4 mmol). The reaction was
stirred at RT for 20 h, partitioned between ethyl acetate (50 mL) and
KHSO.sub.4 solution (50 mL), back-extracted with ethyl acetate (2.times.50
mL), and the combined organic layers were washed with saturated
NaHCO.sub.3 and brine, dried (MgSO.sub.4), filtered and concentrated. The
crude product was chromatographed on silica gel and eluting with 50-67%
EtOAc in hexanes followed by EtOAc to provide title compound (577 mg,
58%).
J.
trans-N-(4-Aminobutyl)-4-(methylthio)-1-(2-naphthalenylsulfonyl)amino!ac
etyl!-L-prolinamide, hydrobromide
The CBZ group was removed from Part I compound (577 mg, 0.96 mmol) by
treating with 30% HBr in HOAc (10 mL) for 1.5 h and then concentrated in
vacuo. Trituration of the residue with ether gave a solid which was
harvested by filtration and washed with more ether leaving the title
deprotected amine (100%).
K.
trans-N-4-(Aminoiminomethyl)amino!butyl!-4-(methylthio)-1-(2-naphthale
nylsulfonyl)amino!acetyl!-L-prolinamide, trifluoroacetate salt
The Part J compound (0.96 mmol) was dissolved in ethanol (20 mL), to which
amidinesulfonic acid (173 mg, 1.4 mmol) and triethyl amine (375 .mu.L, 2.7
mmol) were added. After 4 h the mixture was filtered through a pad of
Celite, washed with ethanol and the filtrate was concentrated to dryness.
The crude material was purified by preparative HPLC (YMC S-10 ODS
50.times.500 mm column, eluting with 54% methanol in water, containing
0.1% TFA). Fractions containing pure title compound were combined and
lyophilized to provide a white solid (279 mg, 44%), Purity.gtoreq.98%.
.alpha.!.sub.D =-12.7.degree., (c=0.7, MeOH).
Analysis calcd for 1.10 TFA+0.90 H.sub.2 O: C, 45.70; H, 5.31; N, 12.69; F,
9.68; S, 9.47. Found: C, 45.65; H, 5.09; N, 12.51; F, 9.74; S, 9.37.
EXAMPLE 7
N-4-(Aminoiminomethyl)amino!butyl!-1-N-(2-naphthalenylsulfonyl)-D-phenyl
alanyl!-L-prolinamide, trifluoroacetate salt, hemihydrate
A. N-4-(1,1-Dimethylethoxy)carbonyl!amino!butyl!-L-prolinamide,
hyrochloride
A(1). (4-Aminobutyl)carbamic acid, 1,1-dimethylethyl ester
To a stirred solution of 1,4-diaminobutane (50 g, 567 mmol) in 195 mL of
dioxane under argon at room temperature was added dropwise a solution of
Di-t-butyl dicarbonate (15.7 g, 71.9 mmol) in 195 mL of dioxane over 3.5
h. Some white precipitate appeared during the addition. The mixture was
stirred at room temperature for 22 h and concentrated in vacuo. The
residue was diluted with 320 mL of water and the precipitate was filtered
off. The aqueous filtrate was extracted with methylene chloride
(3.times.300 mL). The combined methylene chloride extracts were washed
with water (2.times.200 mL) and brine (1.times.200 mL). The organic layer
was dried (MgSO.sub.4), filtered and concentrated in vacuo to give 9.79 g
(72%) of title mono-BOC.amine.
TLC: silica gel, 2% NH.sub.4 OH in 10% CH.sub.3 OH/CH.sub.2 Cl.sub.2,
R.sub.f 0.30, Ninhydrin.
A(2).
N-4-(1,1-Dimethylethoxy)carbonyl!amino!butyl!-1-(phenylmethoxy)carbony
l!-L-prolinamide
To a stirred solution of N-CBZ-L-proline (12.7 g, 50.9 mmol),
1-hydroxybenzotriazole monohydrate (6.49 g, 50.9 mmol) and Part A(1) BOC
amine (9.57 g, 50.9 mmol) in 250 mL of DMF was added in order
4methylmorpholine (11.2 mL, 102 mmol) and ethyl-3-(3-dimethylamino)propyl
carbodiimide hydrochloride (9.76 g, 50.9 mmol). The reaction solution was
stirred at room temperature for 22 h and concentrated under pump vacuum at
50.degree. C. The residue was diluted with 600 mL of EtOAc and washed with
1N HCl solution (2.times.250 mL), saturated NaHCO.sub.3 solution
(2.times.250 mL) and brine (1.times.250 mL). The EtOAC layer was dried
(MgSO.sub.4), filtered and concentrated in vacuo to give 20.7 g (97%) of
title CBZ amine.
TLC: silica gel, 4% CH.sub.3 OH/CH.sub.2 Cl.sub.2, R.sub.f 0.44, UV,
Ce(SO.sub.4).sub.2.
A(3). N-4-(1,1-Dimethylethoxy)carbonyl!-amino!butyl!-L-prolinamide,
hydrochloride
To a stirred solution of Part A(2) CBZ amine (20.2 g, 48.2 mmol) in 250 mL
of methanol under argon was added 20% Pd(OH).sub.2 /C (4.04 g, 20% based
on the weight of Part A(2) compound). The atmosphere was replaced by
hydrogen with several vacuum-fill cycles. The reaction mixture was stirred
at room temperature for 21 h. The catalyst was filtered off through a 4
.mu.M polycarbonate film and rinsed with methanol (3.times.50 mL). The
filtrate was concentrated in vacuo. The oily residue was dissolved in 200
mL of ether and treated with 1N HCl solution in ether (53.0 mL, 53.0
mmol). The solution was concentrated in vacuo. The residue was mixed with
200 mL of toluene and 30 mL of methanol and concentrated in vacuo to give
title amine hydrochloride in quantitative yield (15.5 g) as an oil.
B.
N-4-(1,1-Dimethylethoxy)carbonyl!amino!butyl!-1-N-(phenylmethoxy)carb
onyl!D-phenylalanyl!-L-prolinamide
A stirred solution of N-.alpha.-CBZ-D-phenylalanine (0.56 g, 1.9 mmol) in
6.5 mL of DMF at room temperature under argon was treated with
1-hydroxybenzotriazole (0.29 g, 1.9 mmol) and
ethyl-3-(3dimethylamino)propyl carbodiimide hydrochloride (0.36 g, 1.9
mmol). After 20 minutes, Part A compound was added (0.50 g, 1.6 mmol) and
stirring was carried out for 16 hours. The reaction was quenched by the
addition of 75 mL of 0.25M KHSO.sub.4 solution. The suspension was washed
with EtOAc (2.times.40 mL), the combined EtOAc layers were washed with
0.25M KHSO.sub.4 solution (2.times.40 mL), saturated aqueous KHCO.sub.3
solution (2.times.40 mL), brine, dried (Na.sub.2 SO.sub.4), and
concentrated to yield 1.07 g of a white taffy, which by TLC analysis
appeared to contain unreacted N-.alpha.-CBZ-D-phenylalanine. The crude
product was redissolved in 60 mL of EtOAc, washed with saturated aqueous
KHCO.sub.3 solution (3.times.40 mL), brine, dried (Na.sub.2 SO.sub.4),
concentrated, co-evaporated several times with ether and hexane and
triturated with 50 mL of hexane to yield of title compound (0.78 g, 88%)
as a white solid.
C.
N-4-(1,1-Dimethylethoxy)carbonyl!amino!butyl!-1-D-phenylalanyl-L-prolin
amide
A solution of Part B compound (0.58 g, 1.0 mmol) with 0.12 g of
Pd(OH).sub.2 /carbon in 5.0 mL of methanol was hydrogenated at 1 atm for
3.75 hours. The catalyst was removed by filtration through a porous
membrane (Whatman 0.2.mu. Nylon Autovial), and the filtrate was
concentrated to yield title compound (0.39 g, 88%) as a white, hygroscopic
solid.
D.
N-4-(1,1-Dimethylethoxy)carbonyl!amino!butyl!-1-N-(2-naphthalenylsulfo
nyl)-D-phenylalanyl!-L-prolinamide
Triethylamine (0.25 mL, 1.8 mmol) was added dropwise to a stirred 0.degree.
C. solution of Part C compound (0.39 g, 0.90 mmol) and
2-naphthalenesulfonyl chloride (0.21 g, 0.95 mmol) in 7.0 mL of CH.sub.2
Cl.sub.2. The reaction was carried out at room temperature for 45 minutes.
Dichloromethane was removed under vacuum, the residue was partitioned
between EtOAc (50 mL) and aqueous 0.25M KHSO.sub.4 solution (15 mL), the
EtOAc layer was washed with 0.25M KHSO.sub.4 solution (3.times.15 mL),
brine, dried (Na.sub.2 SO.sub.4), and concentrated to yield 0.61 g Of a
colorless glass. Trituration with 50 mL of hexane yielded title compound
(0.52 g, 93%) as a white solid.
E.
N-(4-Aminobutyl)-1-N-(2-naphthalenylsulfonyl)-D-phenylalanyl!-L-prolinami
de, trifluoroacetate salt
Trifluoroacetic acid (5.0 mL) was added to ice-cooled Part D compound (0.48
g, 0.77 mmol). The reaction solution was stirred at room temperature for
45 minutes. Trifluoroacetic acid was removed under vacuum and
co-evaporated several times with ether and hexane until title compound was
obtained as a white solid (0.51 g, ca. 100%).
F.
N-4-(Aminoiminomethyl)amino!butyl!-1-N-(2-naphthalenylsulfonyl)-D-pheny
lalanyl!-L-prolinamide, trifluoroacetate salt, hemihydrate
A solution of Part E compound (0.49 g, 0.77 mmol) in 6.0 mL of absolute
ethanol was treated with amidine sulfonic acid (0.14 g, 1.2 mmol) followed
by triethylamine (3.2 mL, 2.3 mmol). Within several minutes reaction time,
a precipitate appeared. After 5 hours reaction time, TLC analysis of the
reaction mixture (silica, 3:1:1 nBuOH:HOAc:H.sub.2 O) indicated a small
amount of unconsumed Part E compound. An additional portion of amidine
sulfonic acid (0.05 g) and triethylamine (0.53 mL) were introduced, and
the reaction was continued for 16 hours. TLC analysis as above indicated
complete consumption of Part E compound. The reaction mixture was
concentrated, dissolved in 25 mL of CH.sub.3 OH, and filtered through a
porous membrane (Gelman Acrodisc CR PFTE, 0.45.mu.). Preparative HPLC of
the filtered solution (25% B to 100% B, 30 minute gradient) and subsequent
lyophilization of the pooled fractions yielded title compound, (258 mg,
48%) as a colorless solid. mp 100.degree.-120.degree. C. with foaming.
.alpha.!.sub.D =+19.4.degree. (c=0.5, CH.sub.3 OH).
TLC: Rf=0.70 (silica, 3:1:1 nBuOH; HOAc; H.sub.2 O).
Anal. Calc'd for C.sub.31 H.sub.37 N.sub.6 O.sub.6 F.sub.3 S.1.1 C.sub.2
HF.sub.3 O.sub.2.0.50 H.sub.2 O: C, 53.60; H, 5.49; N, 12.02; F, 8.97; S,
4.59 Found: C, 53.61; H, 5.50; N, 11.81; F, 9.03; S, 4.36
EXAMPLE 8
N-4-(Aminoiminomethyl)
amino!butyl!-1-N-(2-naphthalenylsulfonyl)-D-glutaminyl!-L-prolinamide
A.
N-4-(1,1-Dimethylethoxy)carbonyl!amino!butyl!-1-N-(phenylmethoxy)carb
onyl!D-glutaminyl!-L-prolinamide
To a stirred solution of Example 7 Part A amine hydrochloride (0.56 g, 1.74
mmol), 1-hydroxybenzotriazole monohydrate (0.29 g, 1.74 mmol) and
N-CBZ-D-glutamine (0.49 g, 1.74 mmol) in 15 mL of DMF was added
4-methylmorpholine (0.57 mL, 5.23 mmol) followed by
ethyl-3-(3-dimethylamino)propyl carbodiimide hydrochloride (0.33 g, 1.74
mmol). The solution was stirred at room temperature for 19 h and
concentrated under pump vacuum at 45.degree. C. The oily residue was
diluted with 160 mL of EtOAc and washed with 1N HCl solution (2.times.60
mL), saturated NaHCO.sub.3 solution (2.times.60 mL) and brine (1.times.60
mL). The EtOAc layer was dried (MgSO.sub.4), filtered and concentrated in
vacuo to give 0.75 g (77%) of title CBZ amine.
TLC: silica gel, 6% CH.sub.3 OH/CH.sub.2 Cl.sub.2, R.sub.f 0.22,
Ce(SO.sub.4).sub.2.
B.
N-4-(1,1-Dimethylethoxy)carbonyl!amino!butyl!-1-N-(2-naphthalenylsulfo
nyl)-D-glutaminyl!-L-prolinamide
To a stirred solution of Part A CBZ amine (0.73 g, 1.33 mmol) in 12 mL of
methanol under argon was added Pd(OH).sub.2 /C (146 mg, 20% based on the
weight of Part A compound). The atmosphere was replaced by hydrogen with
several vacuum-fill cycles. The reaction solution was stirred at room
temperature for 22 h and the catalyst was filtered off through a 4 .mu.M
polycarbonate film. The solid was rinsed with methanol (2.times.20 mL).
The filtrate was concentrated in vacuo to give 0.54 g of the intermediate
amine. To a stirred solution of this amine (0.44 g, 1.07 mmol) and
2-naphthalenesulfonyl chloride (0.27 g , 1.18 mmol, in 10 mL of dry
methylene chloride under argon at 0.degree. C. was added Et.sub.3 N (0.33
mL, 2.34 mmol). The reaction solution was stirred at room temperature for
2 h and diluted with 180 mL of EtOAc. This solution was washed with 1N HCl
solution (2.times.60 mL), saturated NaHCO.sub.3 solution (1.times.60 mL)
and brine (1.times.60 mL). The organic layer was dried (MgSO.sub.4),
filtered and concentrated in vacuo. Purification was effected by flash
chromatography on 30 g of Merck silica gel 60 using 240 mL of 4% CH.sub.3
OH/CH.sub.2 Cl.sub.2 and 480 mL of 6% CH.sub.3 OH/CH.sub.2 Cl.sub.2 as
eluants to give pure title sulfonamide (310 mg, 48%).
TLC: silica gel, 4% CH.sub.3 OH/CH.sub.2 Cl.sub.2, R.sub.f 0.20, UV,
I.sub.2.
C.
N-4-(Aminoiminomethyl)amino!butyl!-1-N-(2-naphthalenylsulfonyl)-D-gluta
minyl!-L-prolinamide
To a stirred solution of Part B sulfonamide (310 mg, 0.51 mmol) in 5 mL of
dry methylene chloride was added 0.degree. C. 4N HCl in dioxane (6.00 mL,
24.0 mmol). The solution was stirred at room temperature for 2 h and
diluted with 120 mL of ether. The solid was filtered off and rinsed with
ether (2.times.30 mL). The solid was dissolved in 60 mL of methanol and
concentrated in vacuo to give the intermediate amine hydrochloride. To a
stirred mixture of this amine hydrochloride and Et.sub.3 N (0.39 mL, 2.31
mmol) in 5 mL of absolute EtOH under argon was added
aminoiminomethanesulfonic acid (204 mg, 1.64 mmol). The mixture was
stirred at room temperature for 5 h and concentrated in vacuo. The
reaction mixture was purified by preparative HPLC, eluting isocratically
with a 37% composition of water:methanol (90:10 and 10:90) containing 0.2%
H.sub.3 PO.sub.4. The fractions were concentrated in vacuo and lyophilized
to give 260 mg (93%) of title compound.
m.p. 72.degree.-74.degree. C.
.alpha.!.sub.D =+5.80 (c=0.69, methanol).
Anal. Calc'd for C.sub.25 H.sub.35 N.sub.7 O.sub.5 S.1.60 TFA.1.0 H.sub.2
O: C, 45.40; H, 5.21; N, 13.14; S, 4.30; F, 12.22 Found: C, 45.45; H,
4.98; N, 12.95; S, 4.32; F, 11.90
Following the procedures of Examples 1 to 8 and those outlined above, the
following additional examples may be prepared.
Melting Point .degree.C.,
Optical Rotation and/or
Elemental Analysis (where available)
9.
2R-1(S*),2.alpha.,4.beta.)!-N-3-(Aminoiminomethyl)amino!propyl!-1-4-h
ydroxy-3-(2-naphthalenylsulfonyl)amino)-1,2-dioxobutyl!-2-piperidinecarbox
amide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =+11.0 (C=1.09 MeOH)
Anal. Calc'd for C.sub.24 H.sub.34 N.sub.6 O.sub.5 S.1.1TFA.1.34 H.sub.2 O:
C, 47.10; H, 5.70; N, 12.58; S, 4.80; F, 9.38 Found: C, 47.39; H, 5.41; N,
12.29; S, 5.21; F, 9.56
10.
N-4-(Aminoiminomethyl)amino!butyl!-1-N-(2-naphthalenylsulfonyl)-L-seryl
!-L-prolinamide
.alpha.!.sub.D =-70.2 (c=0.5, MeOH)
Anal. Calc'd for C.sub.23 H.sub.32 N.sub.6 O.sub.5 S.1.15 TFA.0.5 H.sub.2
O: C, 47.13; H, 5.34; N, 13.03; S, 4.97; F, 10.17 Found: C, 46.83; H,
5.28; N, 12.84; S, 5.14; F, 10.43
11.
N-4-(Aminoiminomethyl)amino!butyl!-1-N-(2-naphthalenylsulfonyl)-L-seryl
!-D-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =-14.8 (c=0.5, MeOH)
Anal. Calc'd for C.sub.23 H.sub.32 N.sub.6 O.sub.5 S.1.15 TFA.1.6 H.sub.2
O:
C, 45.73; H, 5.51; N, 12.65; S, 4.82; F, 9.86 Found: C, 45.58; N, 5.19; N,
12.41; S, 5.14; F, 10.01
12.
(2R-trans)-N-4-(Aminoiminomethyl)amino!butyl!-4-methyl-1-(2-naphthalen
ylsulfonyl)amino!acetyl!-2-piperidinecarboxamide, trifluoroacetate salt,
hydrate
.alpha.!.sub.D =+33.0 (c=1.08 MeOH)
Anal. Calc'd for C.sub.24 H.sub.34 N.sub.6 O.sub.4 S.1.35TFA.0.85 H.sub.2
O: C, 47.73; H, 5.56; N, 12.51; S, 4.77; F, 11.45 Found: C, 47.73; H,
5.51; N, 12.13; S, 4.95; F, 11.42
13.
2R-1(S*)2.alpha.,4.beta.!!-N-4-(Aminoiminomethyl)amino!butyl!-1-3-hyd
roxy-2-(2-naphthalenylsulfonyl)amino!-1-oxopropyl!-4-methyl-2-piperidineca
rboxamide, trifluoroacetate salt, hydrate
.alpha.!.sub.D =+18.8 (c=1.06 MeOH)
Anal. Calc'd for C.sub.26 H.sub.38 N.sub.6 O.sub.5 S.1.36TFA.1.00 H.sub.2
O: C, 47.93; H, 5.79; N, 11.68; S, 4.45; F, 10.77 Found: C, 47.92; H,
5.66; N, 11.53; S, 4.89; F, 10.75
14.
1(S)!-N-4-(Aminoiminomethyl)amino!butyl!-1,2,3,4-tetrahydro-2-3-hydrox
y-2-(2-naphthalenylsulfonyl)amino!-1-oxopropyl!-3-isoquinolinecarboxamide,
isomer A, trifluoroacetate salt, hydrate
.alpha.!.sub.D =-30.9 (c=0.5, MeOH)
Anal. Calc'd for C.sub.28 H.sub.34 N.sub.6 O.sub.5 S.1.25 TFA.1.0 H.sub.2
O: C, 50.37; H, 5.16; N, 11.56; S, 4.41; F, 9.80 Found: C, 50.31; H, 5.05;
N, 11.58; S, 4.44; F, 9.65
15.
1(S)!-N-4-(Aminoiminomethyl)amino!butyl!-1,2,3,4-tetrahydro-2-3-hydrox
y-2-(2-naphthalenylsulfonyl)amino!-1-oxopropyl!-3-isoquinolinecarboxamide,
isomer B, trifluoroacetate salt, hydrate
.alpha.!.sub.D =-22.8 (c=0.6, MeOH)
Anal. Calc'd for C.sub.28 H.sub.34 N.sub.6 O.sub.5 S.1.25 TFA.1.0 H.sub.2
O: C, 50.15; H, 5.13; N, 11.47; S, 4.37; F, 10.11 Found: C, 50.04; H,
4.95; N, 11.36; S, 4.51; F, 10.17
16.
(4S*)-N-4-(Aminoiminomethyl)amino!butyl!-4-hydroxy-1-N-(2-naphthalenyls
ulfonyl)-L-seryl!-L-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =-29.0 (c=0.6, MeOH)
Anal. Calc'd for C.sub.23 H.sub.32 N.sub.6 O.sub.6 S.1.15 TFA.1.0 H.sub.2
O: C, 45.25; H, 5.31; N, 12.51; S, 4.77; F, 9.76 Found: C, 45.28; H, 4.98;
N, 12.52; S, 4.86; F, 9.86
17.
(4S*)-N-4-(Aminoiminomethyl)amino!butyl!-4-methoxy-1-N-(2-naphthalenyls
ulfonyl)-L-seryl!-L-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =-24.5 (c=0.6, MeOH)
Anal. Calc'd for C.sub.24 H.sub.34 N.sub.6 O.sub.6 S.1.1 TFA.1.3 H.sub.2 O:
C, 46.04; H, 5.56; N, 12.30; S, 4.69; F, 9.17 Found: C, 46.04; H, 5.48; N,
12.16; S, 4.78; F, 9.22
18.
N-4-(Aminoiminomethyl)amino!butyl!-1-N-(2-naphthalenylsulfonyl)-L-pheny
lalanyl!-L-prolinamide, trifluoroacetate (1:1) salt
m.p. 172.degree.-178.degree. C. with foaming
.alpha.!.sub.D =-27.0 (c=0.5, CH.sub.3 OH)
Anal. Calc'd for C.sub.31 H.sub.37 N.sub.6 O.sub.6 F.sub.3 S.1.2 C.sub.2
HF.sub.3 O.sub.2.0.80 H.sub.2 O: C, 52.68; H, 5.46; N, 11.74; F, 9.55; S,
4.48 Found: C, 52.71; H, 5.35; N, 11.50; F, 9.33; S, 4.26
19.
(2S-trans)-N-4-(Aminoiminomethyl)amino!butyl!-4-methyl-1-(2-naphthalen
ylsulfonyl)amino!acetyl!-2-piperidinecarboxamide
.alpha.!.sub.D =-36.4 (c=0.98, MeOH)
Anal. Cald'd for C.sub.26 H.sub.35 N.sub.6 O.sub.6 S.0.25TFA.1.20 H.sub.2
O: C, 47.74; H, 5.69; N, 12.60; F, 10.68; S, 4.81 Found: C, 47.75; H,
5.51; N, 12.62; F, 10.86; S, 4.95
20.
N-4-(Aminoiminomethyl)amino!butyl!-1-N-(2-naphthalenylsulfonyl)-L-gluta
minyl!-L-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =-48.1 (c=0.68, methanol)
Anal. Calc'd for C.sub.25 H.sub.35 N.sub.7 O.sub.5 S.1.27 TFA.1.57 H.sub.2
O: C, 46.02; H, 5.53; N, 13.64; S, 4.46; F, 10.07 Found: C, 46.02; H,
5.18; N, 13.33; S, 4.86; F, 10.06
21.
N-4-(Aminoiminomethyl)amino!butyl!-1-N-(2-naphthalenylsulfonyl)-L-threo
nyl!-L-prolinamide, trifluoroacetate (1:1) salt
m.p. 68.degree.-78.degree. C.
.alpha.!.sub.D =-72.6 (c=0.69, methanol)
Anal. Calc'd for C.sub.24 H.sub.34 N.sub.6 O.sub.5 S.1.25 TFA. 1.20 H.sub.2
O: C, 46.62; H, 5.56; N, 12.31; S, 4.70; F, 10.43 Found: C, 46.75; H,
5.50; N, 12.08; S, 4.82; F, 10.40
22.
N-4-(Aminoiminomethyl)amino!butyl!-1-N-(2-naphthalenylsulfonyl)-L-allot
hreonyl!-L-prolinamide, trifluoroacetate (1:1) salt
m.p. 85.degree.-96.degree. C.
.alpha.!.sub.D =-70.8 (c=0.76, methanol)
Anal. Calc'd for C.sub.24 H.sub.34 N.sub.6 O.sub.5 S.1.16 TFA.0.97 H.sub.2
O: C, 47.30; H, 5.59; N, 12.57; S, 4.80; F, 9.89 Found: C, 47.30; H, 5.16;
N, 12.39; S, 5.19; F, 9.48
23.
(S)-N-4-(Aminoiminomethyl)amino!butyl!-4-(methylthio)-1-N-(2-naphthalen
ylsulfonyl)glycyl!-L-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =+13.4 (c=0.6, MeOH)
Anal. Calc'd for C.sub.23 H.sub.32 N.sub.6 O.sub.5 S.sub.2.1.10 TFA.1.0
H.sub.2 O: C, 45.58; H, 5.33; N, 12.65; S, 9.44; F, 9.66 Found: C, 45.52;
H, 5.11; N, 12.46; S, 9.49; F, 9.65
24.
(R)-N-4-(Aminoiminomethyl)amino!butyl!-4-(methylthio)-1-N-(2-naphthalen
ylsulfonyl)glycyl!-L-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =-12.7 (c=0.7, MeOH)
Anal. Calc'd for C.sub.23 H.sub.32 N.sub.6 O.sub.4 S.sub.2.1.10 TFA.0.9
H.sub.2 O:
C, 45.70; H, 5.31; N, 12.69; S, 9.47; F, 9.68 Found: C, 45.65; H, 5.09; N,
12.51; S, 9.37; F, 9.74
25.
N-4-(Aminoiminomethyl)amino!butyl!-1-(2-naphthalenylsulfonyl)amino!ace
tyl!-2-piperidinecarboxamide
High Resolution Mass Spectrum: (M+H).sup.+ =489.2287.
26.
N-4-(Aminoiminomethyl)amino!butyl!-1-N-(2-naphthalenylsulfonyl)-D-trypt
ophyl!-L-prolinamide, trifluoroacetate (1 ) salt
m.p.100.degree.-160.degree. C. foam/dec
.alpha.!.sub.D =+44.4 (c=0.5, CH.sub.3 OH)
Anal. Calc'd for C.sub.31 H.sub.37 N.sub.7 O.sub.4 S.1.06 C.sub.2 HF.sub.3
O.sub.2.1.35 H.sub.2 O: C, 53.12; H, 5.49; N, 13.09; F, 8.07; S, 4.28
Found: C, 53.51; H, 5.35; N, 12.73; F, 8.10; S, 4.28
27.
(2S-trans)-N-4-(Aminoiminomethyl)amino!butyl!-4-methyl-1-(3-methyl-8-
quinolinyl)sulfonyl!amino!acetyl!-2-piperidinecarboxamide, trifluoroacetate
salt
.alpha.!.sub.D =-24.1 (c=0.99 MeOH)
Anal. Calc'd for C.sub.24 H.sub.35 N.sub.7 O.sub.4 S.1.37TFA.1.00 H.sub.2
O: C, 46.42; H, 5.59; N, 14:.17; F, 11.29; S, 4.63 Found: C, 46.51; H,
5.32; N, 14.06; F, 11.28; S, 4.55
28.
N-4-(Aminoiminomethyl)amino!butyl!-1-(8-quinolinylsulfonyl)amino!acety
l!-2-piperidinecarboxamide, trifluoroacetate salt
Anal. Calc'd for C.sub.22 H.sub.31 N.sub.7 O.sub.6 S.2C.sub.2 HF.sub.3
O.sub.2.2.20 H.sub.2 O:
C, 41.21; H, 5.04; N, 12.84; F, 14.93; S, 4.20 Found: C, 41.34; H, 4.69; N,
12.39; F, 14.91; S, 4.55
29.
N-4-(Aminoiminomethyl)amino!butyl!-1-N-(2-naphthalenylsulfonyl)-D-.alph
a.-glutamyl!-L-prolinamide, trifluoroacetate (1:1 ) salt
.alpha.!.sub.D =(c=0.60, methanol)
Anal. Calc'd for C.sub.25 H.sub.34 N.sub.6 O.sub.6 S.1.35 TFA.1.13 H.sub.2
O: C, 46.42; H, 5.31; N, 12.16; S, 4.35; F, 10.43 Found: C, 46.42; H,
5.33; N, 11.84; S, 4.66; F, 10.50
EXAMPLE 29
N-4-(Aminoiminomethyl)
amino!butyl!-1-N-(methylsulfonyl)-D-phenylalanyl!-L-prolinamide,
trifluoroacetate (1:1) salt
A. N-4-(Aminoiminomethyl)amino!butyl!-1-D-phenylalanyl-L-prolinamide,
trifluoroacetate salt
A(1).
N-4-(Aminobutyl)!-1-N-(phenylmethoxy)carbonyl!-D-phenylalanyl-L-prolina
mide, trifluoroacetate
The title compound of Example 7, Part B (0.78 g, 1.38 mmol) was dissolved
in trifluoroacetic acid (3.2 mL)). After 3 hours the solvent was
evaporated and then co-evaporated with hexane/ether five times, and dried
16 hours in vacuo to provide a colorless taffy which was used in the next
step.
A(2).
N-4-(Aminoiminomethyl)amino!butyl!-1-N-(phenylmethoxy)carbonyl!-D-phen
ylalanyl-L-prolinamide, trifluoroacetate
The Part A(1) compound (0.80 g, 1.38 mmol) was dissolved in absolute
ethanol (10.9 mL) and treated with amidinesulfonic acid (0.26 g, 1.5 eq),
followed by triethylamine (0.58 mL, 3 eq). After stirring for two hours
the solvent was evaporated and the crude product purified on reverse phase
HPLC to provide a colorless solid (414 mg, 45%).
A(3). N-4-(Aminoiminomethyl)amino!butyl!-1-D-phenylalanyl-L-prolinamide,
trifluoroacetate salt
The title compound of Part A(2) (0.20 g, 0.30 mmol) was dissolved in
methanol (1.5 mL) and hydrogenated at 1 atmosphere and room temperature
for 3 hours with Pd(OH).sub.2 (40 mg of 20% catalyst). The catalyst was
removed by filtration, and the solvent stripped to provide an oil. The oil
was dissolved in methanol, acidified with trifluoroacetic acid (0.2 mL)
and evaporated to dryness, then dissolved in water and lyophilized to
provide the title compound (152 mg).
Optical rotation:=-75.8.degree. (c=0.5, CH.sub.3 OH).
Anal. Calc'd for C.sub.19 H.sub.30 N.sub.6 O.sub.2.2.07 C.sub.2 HF.sub.3
O.sub.2.0.90 H.sub.2 O): C, 44.35; H, 5.45; N, 13.41; F, 18.82 Found: C,
44.52; H, 5.01; N, 13.26; F, 18.52.
B.
N-4-(Aminoiminomethyl)amino!butyl!-1-N-(methylsulfonyl)-D-phenylalanyl!
-L-prolinamide, trifluoroacetate (1:1) salt
To a stirred solution of Part A amine (550 mg, 0.80 mmol) in 15 mL of dry
CH.sub.2 Cl.sub.2 and 15 mL of dry THF under argon was added in order
triethylamine (Et.sub.3 N) (0.44 mL, 3.20 mmol) and methanesulfonyl
chloride (68.0 .mu.L, 0.88 mmol). This turbid mixture was stirred at room
temperature for 3 h and diluted with 0.50 mL of water. The mixture was
stirred at room temperature for 10 min and concentrated in vacuo. The
residue was diluted with 30 mL of methanol, concentrated in vacuo and
purified by preparative HPLC. The fractions were concentrated in vacuo and
lyophilized to give 300 mg (75%) of title compound.
Opt. rot.: .alpha.!.sub.D =-68.7.degree. (c=1.00, methanol).
Elemental Analysis (%) Calc'd: C, 43.59; H, 5.31; N, 13.09; S, 4.99; F,
14.65 Found: C, 43.59; H, 5.33; N, 13.08; S, 4.80; F, 14.69
EXAMPLE 30
##STR73##
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(sulfonyl)-D-phenylalan
yl!-L-prolinamide
A.
4-(Aminomethyl)-N,N'-bis(1,1-dimethylethoxy)carbonyl!-1-piperidinecarboxi
midamide
To a stirred solution of 4-aminomethylpiperidine (0.72 g, 6.31 mmol) in 40
mL of toluene was added benzaldehyde (0.78 mL, 6.94 mmol). The reaction
solution was refluxed for 18 h and water was removed by a Dean Stark trap.
The reaction solution was cooled to room temperature at which time bis-Boc
amidinopyrazole (1.96 g, 6.31 mmol) was added. The reaction solution was
stirred at room temperature for 48 h and concentrated in vacuo. The oily
residue was diluted with 15 mL of 1M (aq) KHSO4 solution and stirred at
room temperature for 5 h. This aqueous solution was washed with ether
(2.times.20 mL) and basified to pH 12 by the addition of 1N NaOH solution.
This basic solution was then saturated with NaCl and extracted with
dichloromethane (3.times.60 mL). The combined dichloromethane extracts
were dried (MgSO.sub.4), filtered and concentrated in vacuo to give 2.10 g
(93%) of title amine which was used for the next transformation without
further purification.
B.
(S*)-N-1-(1,1-Dimethylethoxy)carbonyl!amino!(1,1-dimethylethoxy)car
bonyl!imino!methyl!-4-piperidinyl!methyl!-1-1-(1,1-dimethylethoxy)carbon
yl!amino!-2-phenylethyl!-L-prolinamide
To a stirred solution of N-Boc-D-Phe-L-Pro-OH (0.73 g, 2.02 mmol), Part A
amine (0.72 g, 2.02 mmol) and 1-hydroxybenzotriazole monohydrate (0.34 g,
2.02 mmol) in 30 mL of DMF was added in order 4-methylmorpholine (0.66 mL,
6.05 mmol) and ethyl-3-(3dimethylamino)propyl carbodiimide hydrochloride
(0.39 g, 2.02 mmol). The reaction solution was stirred at room temperature
for 19 h and concentrated under pump vacuum at 45.degree. C. The residue
was diluted with 100 mL of saturated NaHCO.sub.3 solution and extracted
with dichloromethane (4.times.100 mL). The combined dichloromethane
extracts were dried (MgSO.sub.4), filtered and concentrated in vacuo. This
was chromatographed on silica gel to give 0.70 g (50%) of title bis-Boc
guanidine.
C.
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-D-phenylalanyl-L-prolinam
ide
To a stirred solution of Part B bis-Boc guanidine (0.68 g, 0.97 mmol) in
6.0 mL of dichloromethane was added trifluoroacetic acid (TFA) (6.00 mL,
77.9 mmol). The reaction solution was stirred at room temperature for 3 h
and concentrated in vacuo. This was purified by prep HPLC to give 310 mg
(45%) of title compound.
D.
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(methylsulfonyl)-D-phe
nylalanyl!-L-prolinamide
To a stirred solution of Part C guanidine (275 mg, 0.68 mmol) in 4.0 mL of
dichloromethane and 2.0 mL of THF under argon at 0.degree. C. was added
Et.sub.3 N (0.38 mL, 2.74 mmol) and methanesulfonyl chloride (69 .mu.L) in
order. The reaction mixture was stirred at room temperature for 2.5 h and
diluted with 2.0 mL of water. The mixture was concentrated in vacuo, and
purified by prep HPLC to give 160 mg (37%) of title compound.
EXAMPLE 30a
Alternate Route for Preparation of
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(methylsulfonyl)-D-phe
nylalanyl!-L-prolinamide
To a suspension of N-methylsulfonyl-D-phenylalanyl-proline (497 mg, 1.46
mmol), 4-(aminomethyl)-1-piperidinecarboximidamide (470 mg, 1.9 mmol) and
HOBt (217 mg, 1.6 mmol) in 1:1 v/v isopropyl alcohol/water (4.5 mL) was
added NMM (0.32 mL, 2.9 mmol). The homogeneous solution was cooled to
0.degree. C. and
ethyl-3-(3-dimethylamino)-propylcarbodiimide.hydrochloride salt (EDAC.HCl)
(308 mg, 1.6 mmol) was added. After 18 hr the reaction was concentrated in
vacuo, dissolved in water (10 mL) and washed with ether (2.times.75 mL).
Aqueous NaOH (1N, 3.8 mL) was added until the aqueous layer attained pH
9.7, the aqueous layer was washed with ether (75 mL) and the process
repeated. Aqueous HCl (1N, 3.8 mL) was added until the pH reached 2.0, the
aqueous layer was washed with ethyl acetate (3.times.75 mL), concentrated
in vacuo, dissolved in water, and lyophilized to give a foam which was
subjected to chromatography on HP-20 polystyrene resin to provide the
title compound (629 mg, 84%).
EXAMPLE 30b
N-(1-Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(methylsulfonyl)-D-phen
ylalanyl!-L-prolinamide, acetate (1:1)
##STR74##
A. 4-(Aminomethyl)-1-piperidinyl!iminomethyl!carbamic acid, phenylmethyl
ester
##STR75##
A 500-mL, round-bottomed flask equipped with Dean-Stark trap was
sequentially charged with 4-aminomethylpiperidine (14.84 g,. 129.96
mmole), toluene (250 mL), and benzaldehyde (14.98 g, 141.16 mmole). The
mixture was heated to reflux under an argon atmosphere and stirred
overnight (18 h). The reaction was cooled to room temperature and the
toluene was removed in vacuo. Residual toluene was removed under high
vaccuum to provide 28.14 g of the title compound as an orange oil
(quantitative yield). The crude product was carried directly into the next
step.
##STR76##
A 500-mL, round-bottomed flask was sequentially charged with Part A(1)
Schiff base (21.20 g, 104.8 mmole), acetonitrile (200 mL),
imino(1H-pyrazol-1-yl)methyl!carbamic acid, phenylmethyl ester (31.50 g,
128.96 mmole) and 1,8diazabicyclo5.4.0!undec-7-ene (DBU) (18 mL, 120.36
mmole). After stirring for five minutes, the solution became clear yellow.
The reaction was heated to 50.degree. C. and stirred under a nitrogen
atmosphere for 16 h. After the reaction had cooled to room temperature,
12N hydrochloric acid (36 mL, 432 mole, reaction mixture reached pH 1) was
added over 20 minutes (exotherm to 40.degree. C.). The reaction was
stirred for 2.5 h at room temperature. The mixture was concentrated in
vacuo to a yellow oil and isopropanol (35 mL) was added. With stirring,
acetonitrile (250 mL) was added resulting in the separation of an oil. The
mixture was heated to 40.degree. C. providing a clear solution which was
seeded. Within five minutes a white solid crystallized from solution. The
slurry was stirred for an additional 40 minutes at 40.degree. C. and then
cooled to room temperature and stirred overnight. The product was
collected by filtration and washed with 1:1 ethyl acetate/hexane
(1.times.100 mL), 1:3 ethyl acetate/hexane (1.times.100 mL), and hexane
(3.times.100 mL). Residual solvents were removed under high vaccuum to
provide 29.95 g of an 11:1 mixture of
imino(1H-pyrazol-1-yl)methyl!carbamic acid, phenylmethyl ester,
dihydrochloride salt:4-aminomethyl piperidine, dihydrochloride salt as a
white solid (79%).
A(3). 4-(Aminomethyl)-1-piperidinyl!iminomethyl!carbamic acid,
phenylmethyl ester
Sodium hydroxide (4.45 g, 111.25 mmole) was dissolved in 150 mL of
deionized water. The mixture of imino(1H-pyrazol-1-yl)methyl!carbamic
acid, phenylmethyl ester, dihydrochloride salt and 4-aminomethyl
piperidine, dihydrochloride salt (prepared above, 16.18 g, 44.54 mmole)
was added (pH of the cloudy solution was 13) and the mixture was extracted
with dichloromethane (1.times.200 mL and 2.times.100 mL). The organic
layers were combined and washed with brine (1.times.200 mL), dried
(MgSO.sub.4), filtered and concentrated in vacuo to provide the title
compound as a white foam (12.29 g, 95%).
B. N-(Methylsulfonyl)-D-phenylalanyl!-L-proline
##STR77##
D-Phenylalanine (15.01 g, 90.865 mmol) was weighed into a 250-mL, 5-necked,
round-bottomed flask equipped with thermocouple, pH probe, overhead
stirrer and two addition funnels (one a constant-rate funnel to ensure
even addition of methanesulfonyl chloride). Sodium hydroxide (3.642 g,
91.05 mmol) was dissolved in 90 mL of deionized water and added to the
reaction flask. With stirring, the mixture became a clear solution. The
internal temperature was brought to -2.degree. C. (pH 12.3). With rapid
stirring, methanesulfonyl chloride (5.0 mL, 64.60 mmol) was added via the
constant-rate addition funnel. Once pH 11 was reached, 6N NaOH was added
dropwise to maintain the pH at 10.9.+-.0.2. A second portion of
methanesulfonyl chloride (4.2 mL, 54.26 mmol) was then added while
maintaining the pH at 10.5.+-.0.2 (internal temperature <8.degree.) by
dropwise addition of 6N sodium hydroxide. Once the pH had stabilized at
10.5, HPLC anlysis of an aliquot showed a 9:1 mixture of the title
compound to D-phenylalanine. A third portion of methanesulfonyl chloride
(1.2 mL, 15.5 mmol) was then added at 0.degree. over 5 minutes while
maintaining the pH at 10.5.+-.0.2. After the addition was complete and the
pH had stabilized at 10.4, the reaction mixture was brought to pH 13 with
6N sodium hydroxide, allowed to warm to 15 .degree. C. over 2 hr, and then
washed with methyl isobutylketone (2.times.100 mL). The aqueous layer was
acidified to pH 1 with 6N hydrochloric acid and extracted with ethyl
acetate (1.times.200 mL and 2.times.150 mL). The ethyl acetate layers were
combined and washed with brine (1.times.150 mL), dried over MgSO.sub.4,
and concentrated in vacuo to yield a yellow oil (18.36 g, 83%). HPLC
analysis of the oil showed a 7:1 mixture of the title compound to
N-methylsulfonyl-D-phenylalanine!-D-phenylalanine. This material was
carried into the next step without further purification.
##STR78##
The Part B(1) mixture of N-methylsulfonyl-D-phenylalanine and
N-methylsulfonyl-D-phenylalanine!-D-phenylalanine (18.36 g, 75.47 mmol)
was dissolved in 250 mL of dichloromethane under an argon atmosphere.
Heating to reflux and stirring for about 15 minutes was necessary to
completely dissolve the material. After the solution had cooled to room
temperature, DMF (0.23 mL, 2.97 mmol) was added and the internal
temperature was further lowered to 3 .degree. C. (ice bath). Oxalyl
chloride (7.4 mL, 84.85 mmol) was added in a steady stream over 2 minutes.
After the addition was complete, the reaction was allowed to warm to room
temperature over 3 hours and then stirred at room temperature for one
hour. The mixture was concentrated in vacuo to a yellow oil, toluene (100
mL) was added and the solution was concentrated to a yellow oily solid
(19.89 g, 101%). This material was stored in a refrigerator overnight
under an argon atmosphere and was carried into the next step without
further purification.
B(3). N-(Methylsulfonyl)-D-phenylalanyl!-L-proline
The crude Part B(2) acid chloride prepared above was dissolved in 120 mL of
toluene, with heating to 40.degree. C., and then cooled to room
temperature under an argon atmosphere. L-proline (13.11 g, 113.87 mmol)
was weighed into a 500-mL, 5-necked flask equipped with two addition
funnels, thermocouple, pH probe and overhead stirrer. Sodium hydroxide
(2.68 g, 67 mmol) was dissolved in 120 mL of deionized water and added to
the reaction flask. The proline readily dissolved with stirring to give a
clear solution which was cooled to 0.degree. C. The initial pH was 12.15.
The acid chloride-toluene solution was added dropwise to the rapidly
stirring proline solution until the pH reached 11.0 (internal temperature
maintained at <5.degree. C.). The remainder of the acid chloride was added
dropwise over 25 minutes while maintaining the internal temperature below
5.degree. C. and the pH at 11.0.+-.0.2 by dropwise addition of 4N sodium
hydroxide (23 mL total). The acid chloride was washed in with 20 mL of
toluene. After the addition was complete and the pH had stabilized to
11.1, the icebath was removed and the reaction was allowed to warm to
12.degree. C. over 30 minutes. The aqueous and organic layers were
separated and the basic aqueous layer was washed with ethyl acetate
(1.times.100 mL, discard). Ethyl acetate (350 mL) was added to the basic
aqueous solution and while stirring vigorously, the mixture was acidified
with 6N hydrochloric acid to pH 1. The organic and aqueous layers were
separated and the acidic aqueous was extracted with ethyl acetate
(2.times.150 mL). The combined ethyl acetate layers from the acidic
extractions were washed with brine (1.times.150 mL), dried (MgSO.sub.4),
filtered, and concentrated in vacuo to a total weight of 260 g, providing
a slurry of the title compound in the ethyl acetate. With stirring, 260 mL
of hexane was added and the slurry was stirred for 4 h at room
temperature. The white crystalline solid was collected by filtration,
washed with hexane/ethyl acetate (2:1, 2.times.100 mL) followed by hexane
(2.times.100 mL). Drying in vacuo provided the title compound (20.84 g,
84%) as a white crystalline solid.
C.
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(methylsulfonyl)-D-phe
nylalanyl!-L-prolinamide, acetate (1:1)
##STR79##
A 500-mL, round-bottomed flask was sequentially charged with Part A
compound (10.41 g, 35.84 mmole), isopropanol (60 mL), deionized water (60
mL), and Part B compound (10.00 g, 29.38 mmole). Under an argon
atmosphere, the clear yellow solution was brought to 10.degree. C. and
1-hydroxybenzotriazole hydrate (4.40 g, 32.56 mmole) was added. The
mixture was further cooled to 4.degree. C. and
1-(3-dimethylaminopropyl)-3-ethlycarbodiimide hydrochloride (6.53 g, 34.06
mole) was added. The resulting clear solution was stirred between
1.degree. and 4.degree. C. for 3 hours and then slowly warmed to
20.degree. C. over 14 hours. A portion of the solvent (62 g) was removed
in vacuo. Ethyl acetate (250 mL) was added. With stirring, deionized water
(465 mL) and 6N hydrochloric acid (40 mL) were added. The aqueous and
organic layers were separated and the aqueous layer was washed with ethyl
acetate (1.times.100 mL). The combined ethyl acetate layers were
concentrated to 100 g and extracted with 0.5N hydrochloric acid
(2.times.60 mL). The combined acidic aqueous layers were brought to pH 6
with 12N sodium hydroxide and extracted with dichloromethane (1.times.250
mL and 2.times.150 mL). The dichloromethane layers were combined and
washed with deionized water (1.times.150 mL) and brine (1.times.150 mL),
dried (MgSO.sub.4), filtered and concentrated in vacuo. The residual
solvents were removed under high vacuum to provide title CBZ-protected
compound as a white foam (15.44 g, 83% yield, corrected for ethyl acetate
in .sup.1 H NMR). The crude product was carried directly into the next
step.
C(2).
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(methylsulfonyl)-D-phe
nylalanyl!-L-prolinamide, acetate (1:1)
A solution of Part C(1) CBZ-protected compound (13.22 g, 21.58 mole) in
MeOH (150 mL) was purged with argon for ten minutes. Palladium hydroxide
on carbon (1.32 g) was added and the mixture was again purged with argon
for ten minutes. The solution was then purged with hydrogen and the
reaction was stirred under a hydrogen atmosphere for 2.5 hours. After the
reaction was complete by HPLC, the mixture was purged with argon, filtered
through celite and the celite washed with methanol (2.times.100 mL).
Acetic acid was added and the solution was concentrated in vacuo to an
oil. Isopropanol (100 mL) was added and the solution concentrated in vacuo
to 45 g. The thick solution was seeded. With stirring, 180 mL of
acetonitrile was added over 20 minutes. Within three hours, a white solid
crystallized from solution and the resulting slurry was allowed to stir
overnight at room temperature. The solid was collected by filtration and
washed with acetonitrile (1.times.50 mL), 1:1 hexane/ethyl acetate
(1.times.60 mL), and hexane (2.times.50 mL). The product was dried in
vacuo to yield the title compound as a white crystalline solid (8.83 g,
76%).
EXAMPLE 31
N-1-(Aminoiminomethyl)-3-piperidinyl!methyl!-1-N-(methylsulfonyl)-D-phen
ylalanyl!-L-prolinamide
A. N-Boc-3-hydroxymethylpiperidine
To a stirred solution of 3-hydroxymethyl-piperidine (15.1 g, 131 mmol) and
Et.sub.3 N (21.9 mL, 158 mmol) in 100 mL of dichloromethane was added
dropwise a solution of di-t-butyl dicarbonate (31.5 g, 144 mmol) in 100 mL
of dichloromethane over 1 h. The reaction was stirred at room temperature
for 18 h and then diluted with 200 mL of dichloromethane. The resulting
solution was washed with 1N HCl solution (3.times.100 mL), saturated
NaHCO.sub.3 solution (2.times.100 mL) and brine (1.times.100 mL). The
organic layer was dried (MgSO.sub.4), filtered and concentrated in vacuo
to give N-Boc-3-hydroxymethylpiperidine (27.0 g, 96%).
B. 3-(Azidomethyl)-1-piperidinecarboxylic acid, 1,1-dimethylethyl ester
To a stirred solution of Part A N-Boc-3-hydroxymethylpiperidine (27.0 g,
126 mmol) in 150 mL of dry dichloromethane under argon at 0.degree. C. was
added in order triethylamine (22.7 mL, 163 mmol) and methanesulfonyl
chloride (11.7 mL, 151 mmol). The reaction was stirred at room temperature
for 1.5 h and diluted with 450 mL of dichloromethane. The reaction was
washed with 0.degree. C. 1N HCl solution (2.times.100 mL) and brine
(1.times.100 mL). The dichloromethane layer was dried (Na.sub.2 SO.sub.4),
filtered and concentrated in vacuo. The residue was dissolved in 200 mL of
DMF and combined with sodium azide (24.5 g, 377 mmol). The mixture was
stirred at room temperature for 33 h and the solid was filtered off. The
filtrate was concentrated under pump vacuum at 45.degree. C. The residue
was partitioned between 400 mL of EtOAc and 10% sodium thiosulfate
solution (2.times.100 mL) and brine (1.times.100 mL). The EtOAc layer was
dried (MgSO.sub.4), filtered and concentrated in vacuo. Purification was
effected by a flash column chromatography on silica gel to give 19.5 g
(65%) of title azide.
C. 3-(Aminomethyl)-1-piperidinecarboxylic acid, 1,1-dimethylethyl ester
To a stirred solution of Part B azide (19.0 g, 79.2 mmol) in 250 mL of
methanol under argon was added 10% Pd/C (3.80 g, 20% based on the weight
of Part B azide). The atmosphere was replaced with hydrogen by several
vacuum-fill cycles. The mixture was stirred at room temperature for 15 h.
The catalyst was filtered through a 4 .mu.M polycarbonate film and rinsed
with methanol (4.times.30 mL). The filtrate was concentrated in vacuo to
give 16.3 g (96%) of title amine.
D.
N-1-(1,1-Dimethylethoxy)carbonyl!-3-piperidinyl!methyl!-1-N-(phenylme
thoxy)carbonyl!-D-phenylalanyl!-L-prolinamide
To a stirred solution of Part C amine (2.00 g, 9,35 mmol),
N-Cbz-D-Phe-L-Pro (3.70 g, 9.35 mmol), 1-hydroxybenzotriazole monohydrate
(1.58 g, 9.35 mmol) and 4-methylmorpholine (3.07 mL, 28.0 mmol) was added
ethyl-3-(3-dimethylamino)propyl carbodiimide hydrochloride (1.79 g, 9.35
mmol). The reaction solution was stirred at room temperature for 17 h and
concentrated under pump vacuum at 45.degree. C. The residue was dissolved
in 360 mL of EtOAc and washed with 1N HCl solution (2.times.120 mL),
saturated NaHCO.sub.3 solution (1.times.120 mL) and brine (1.times.120
mL). The EtOAc layer was dried (MgSO.sub.4), filtered, concentrated in
vacuo and chromatographed on silica gel to give 1.30 g (23%) of title
carbamate.
E.
N-1-(Aminoiminomethyl)-3-piperidinyl!methyl!-1-N-(phenylmethoxy)carbon
yl!-D-phenylalanyl!-L-prolinamide
To a stirred solution of Part D carbamate (2.30 g, 3.89 mmol) in 10 mL of
dry dichloromethane was added 0.degree. C. 4N HCl in dioxane (15.0 mL,
60.0 mmol). The solution was stirred at room temperature for 3 h and
diluted with 300 mL of ether. The precipitate was filtered off and rinsed
with ether (3.times.30 mL). The precipitate was dried under pump vacuum at
room temperature and purified by prep HPLC to give 1.39 g (59%) of
intermediate amine.TFA salt. To a stirred solution of the intermediate
amine.TFA salt (500 mg, 0.83 mmol) and diisopropylethyl amine (0.35 mL,
1.98 mmol) in 2.0 mL of DMF was added 1H-pyrazole-1-carboxamidine (133 mg,
0.91 mmol). The reaction solution was stirred at room temperature for 6 h
and diluted with 100 mL of ether. The desired oily precipitate was
separated from the ether solution and purified by prep HPLC to give 250 mg
(47%) of title Cbz-carbamate.
F.
N-1-(Aminoiminomethyl)-3-piperidinyl!methyl!-1-D-phenylalanyl-L-prolinam
ide
To a stirred solution of Part E Cbz-carbamate (240 mg, 0.37 mmol) in 10 mL
of methanol under argon was added 20% Pd(OH).sub.2 /C (48 mg, 20% based on
the weight of Part E Cbz-carbamate). The atmosphere was replaced with
hydrogen by several vacuum-fill cycles. The reaction mixture was stirred
at room temperature for 24 h. The catalyst was filtered off and rinsed
with methanol (4.times.20 mL). The filtrate was concentrated in vacuo. The
residue was dissolved in 50 mL of a solution of 0.1% TFA in water and
lyophilized to give 220 mg (82%) of title compound.
G.
N-1-(Aminoiminomethyl)-3-piperidinyl!methyl!-1-N-(methylsulfonyl)-D-phe
nylalanyl!-L-prolinamide
Following the procedure of Example 30 Part D except substituting the Part F
amidine for the Example 30 Part C amidine, the title compound is obtained.
EXAMPLE 32
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(phenylmethyl)sulfonyl
!-D-alanyl!-L-prolinamide, trifluoroacetate (1:1) salt
A. N-(Phenylmethoxy)-carbonyl!-D-alanyl!-L-proline methyl ester
To a solution of N-CBZ-D-alanine (28.1 g, 0.149 mol) in DMF (250 mL) at
0.degree. C., was added L-proline-methylester.HCl (24.6 g, 0.149 mmol),
1-hydroxybenzotriazole hydrate (22.2 g, 0.164 mmol),
ethyl-3-(3-dimethylamino)-propyl carbodiimide.HCl (31.4 g, 0.164 mmol) and
4-methylmorpholine (22.6 g, 0.224 mmol). The reaction mixture was stirred
for 12 h while allowing the reaction to warm to room temperature. The
reaction mixture was poured in water (750 mL) and extracted with ethyl
acetate (2.times.150 mL). The combined organic extracts were washed as
follows: KHSO.sub.4 (0.25M, 2.times.50 mL), water (1.times.50 mL),
saturated aqueous NaHCO.sub.3 (2.times.50 mL) and saturated NaCl
(1.times.50 mL). The organic layer was dried over MgSO.sub.4, filtered and
the solvent removed in vacuo to give the title compound, 41.16 g (92%), as
an oil. MS (M+H).sup.+ =301.sup.+.
B. N-(Phenylmethyl)sulfonyl!carbonyl!-D-alanyl!-L-proline methyl ester
A solution of Part A compound (1.56 g, 5.2 mmol) in trifluoroacetic acid (3
mL) was stirred at 0.degree. C. for 1.5 hr and and concentrated in vacuo
to give the corresponding TFA salt. This salt was dissolved in chloroform
(10 mL), and triethylamine (2.2 mL, 15.6 mmol) and benzylsulfonyl chloride
(1.48, 7.8 mmol) at 0.degree. C. were added to it. The reaction mixture
was stirred to room temperature overnight. After 25 h, the reaction
mixture was poured into EtOAc (50 mL), washed with 0.25M aqueous
KHSO.sub.4 (2.times.20 mL), H.sub.2 O (1.times.20 mL) and saturated
aqueous NaHCO.sub.3 (2.times.20 mL), successively; dried over MgSO.sub.4,
filtered and concentrated in vacuo to give title compound as an oil (1.56
g, 85%): MS (M+H).sup.+ =355.sup.+.
C. N-(Phenylmethyl)sulfonyl!carbonyl!-D-alanyl!-L-proline
To a solution of Part B compound (1.33 g, 3.75 mmole) in THF (10 mL) at
0.degree. C. was added aqueous NaOH (1.0N, 9.5 mL). After 30 min the
reaction mixture was warmed to room temperature and stirring continued for
an additional 4 h. The organic solvent was removed in vacuo. The aqueous
layer was acidified to pH of 2-3 and extracted with ethyl acetate
(3.times.10 mL). The combined extracts were dried over MgSO.sub.4,
filtered and the solvent removed in vacuo to give title compound, 1.21 g
(94%), as a white solid. MS (M+H).sup.+ =341.sup.+.
D.
N-1-(1,1-Dimethylethoxy)carbonyl!-4-piperidinyl!methyl!-1-N-(phenylme
thyl)sulfonyl!-D-alanyl!-L-prolinamide
To a solution of Part C compound (1.09 g, 3.2 mmol) in DMF (15 mL) at
0.degree. C. were added
N-1,1(dimethylethoxy)carboxyl!-4-methylaminopiperidine (prepared in
Example 33 Part A) (0.685 g, 3.2 mmol), 1-hydroxybenzotriazole hydrate
(0.476 g, 3.5 mmol), ethyl-3-(3-dimethylamino)-propyl carbodiimide.HCl
(0.675 g, 3.5 mmol) and 4-methylmorpholine untill basic by pH paper. The
reaction mixture was stirred for 12 h while allowing the reaction to warm
to room temperature. The reaction mixture was poured into 0.25M aqueous
KHSO.sub.4 (50 mL) and extracted with EtOAc (2.times.20 mL). The organic
layers were combined and washed with 0.25M aqueous KHSO.sub.4 (1.times.30
mL), water (1.times.25 mL), saturated aqueous NaHCO.sub.3 (2.times.25 mL)
and water (1.times.20 mL). The organic layer was dried over MgSO.sub.4,
filtered and the solvent removed in vacuo to give title compound, 1.67 g
(9%), as an oil. MS (M+H).sup.+ =537.sup.+.
E.
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(phenylmethyl)sulfony
l!-D-alanyl!-L-prolinamide, trifluoroacetate (1:1) salt
A solution of Part D compound (1.17 g, 2.2 mmol) in trifluoroacetic acid
(10.0 mL) was stirred at 0.degree. C. for 5 h. The reaction mixture was
concentrated in vacuo to give the corresponding TFA salt. To a solution of
this salt (2.2 mmol), 1H-pyrazole-1-carboxamidine hydrochloride (0.474 g,
3.3 mmol) and N,N-diisopropylethylamine (1.1 mL, 6.5 mmol) in DMF (3.0 mL)
were added and stirred for 3 days. The reaction mixture was concentrated
in vacuo and purified by preparative HPLC (CH.sub.3 CN/H.sub.2 O with TFA
buffer using a C-18 silica gel column). The appropriate fractions were
combined, concentrated in vacuo, dissolved in H.sub.2 O and lyophilized to
yield title compound (0.486 g, 38%) as a white solid:
a!.sub.D =+1.4 (C 1.0, MeOH)
MS (M+H).sup.+ =479.sup.+
Anal. Calc'd for C22H.sub.34 N.sub.6 O.sub.4 S.1.3 TFA.0.7 H.sub.2 O: C,
46.21; H, 5.78; N, 13.14; S, 5.01; F, 11.59 Found: C, 46.42; H, 5.83; N,
12.88; S, 4.97; F, 11.82
EXAMPLE 33
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(2,2,2-trifluoroethyl)
sulfonyl!-D-phenylalanyl!-L-prolinamide, trifluoroacetate (1:1) salt
A. N-1,1-(Dimethylethoxy)carbonyl!-4-methylamino piperidine
Benzaldehyde (14.6 g, 0.138 mmol), 4-(aminomethyl)-piperdine (14.29 g,
0.125 mmol) and toluene (250 mL) were combined and heated at reflux for
4.5 h with removal of water. The reaction mixture was cooled to
-25.degree. C. and di-t-butyl dicarbonate (28.7 g, 0.131) was added. The
reaction mixture was allowed to warm to room temperature and stirred an
additional 8 h. To this reaction mixture was added aqueous KHSO.sub.4
(1.0M, 120 mL) and stirred for 3 h. The organic layer was removed and the
remaining aqueous layer was extracted with ether (3.times.75 mL). The
aqueous layer was made basic through the addition of aqueous NaOH (1.0M,
130 mL) and extracted with ether (3.times.75 mL). The combined organic
extracts were dried over NaSO.sub.4 filtered, and the solvent removed in
vacuo to give title compound, 24.4 g (9%), as a white solid.
B. N-(phenylmethoxy)-carbonyl!-D-phenyl-alanyl!-L-proline methyl ester
To a solution of N-CBZ-D-phenylalanine (47.3 g, 0.158 mmol) in DMF (300 mL)
at 0.degree. C., was added L-proline-methylester.HCl (25.0 g, 0.151 mmol),
1-hydroxybenzotriazole hydrate (20.28 g, 0.166 mmol),
ethyl-3-(3-dimethylamino)-propyl carbodiimide.HCl (31.8 g, 0.166 mmol) and
4-methylmorpholine (16.0 g, 0.159 mmol). The reaction mixture was stirred
for 8 h while allowing the reaction to warm to room temperature. The
reaction mixture was poured in water (1.2 L) and extracted with ethyl
acetate (3.times.300 mL). The combined organic extracts were washed as
follows: KHSO.sub.4 (0.25M, 2.times.200 mL), water (1.times.200 mL),
saturated aqueous NaHCO.sub.3 (2.times.200 mL) and saturated NaCl
(1.times.200 mL). The organic layer was dried over MgSO.sub.4, filtered
and the solvent removed in vacuo to give title compound, 60.9 g (98%), as
an oil. MS (M+H).sup.+ =411.sup.+.
C. N-(Phenylmethoxy)-carbonyl!-D-phenyl-alanyl-L-proline
To a solution of Part B compound (57.8 g, 0.141 mol) in MeOH (180 mL) and
THF (60 mL) at 0.degree. C. was added aqueous NaOH (1.0N, 183 mL). After
30 min the reaction mixture was warmed to room temperature and stirring
continued for an additional 4 h. Aqueous HCl (1.0N, 42 mL) was added and
the organic solvents removed in vacuo. The resulting aqueous layer was
acidified to pH of 2-3 and extracted with ethyl acetate (2.times.300 mL).
The combined extracts were dried over MgSO.sub.4, filtered and the solvent
removed in vacuo to give title compound, 45.53 g (97%), as a white solid.
MS (M+H).sup.+ =397.sup.+.
D.
N-1-(1,1-Dimethylethoxy)carbonyl!-4-piperidinyl!methyl!-1-N-(phenylme
thoxy)carbonyl!-D-phenylalanyl!-L-prolinamide
To a solution of Part C compound (19.39 g, 48.90 mmol) in DMF (120 mL) at
0.degree. C., was added Part A compound (9.98 g, 46.57 mmol),
1-hydroxybenzotriazole hydrate (6.26 g, 51.23 mmol) and
ethyl-3-(3-dimethylamino)-propyl carbodiimide.HCl (9.82 g, 51.23 mmol).
The reaction mixture was stirred for 12 h while allowing the reaction to
warm to room temperature. The reaction mixture was poured in water (500
mL) and extracted with ethyl acetate (3.times.100 mL). The combined
organic extracts were washed as follows: KHSO.sub.4 (0.25M, 2.times.50
mL), water (1.times.75 mL), saturated aqueous NaHCO.sub.3 (2.times.75 mL)
and saturated NaCl (1.times.50 mL). The organic layer was dried over
MgSO.sub.4, filtered and the solvent removed in vacuo to give title
compound, 23.4 g (85%), as an oil. MS (M+H).sup.+ =593.sup.+.
E.
N-1-(1,1-Dimethylethoxy)carbonyl!-4-piperidinyl!methyl!-1-D-phenylalan
yl!-L-prolinamide
To a stirred solution of Part D compound (15.00 g, 25.3 mmol) in MeOH (150
mL) was added Pd/C (1.50 g) and the reaction mixture was placed under 1
atmosphere of hydrogen. An additional Pd/C (2.5 g) was added after
stirring for 2 h. The reaction was complete after stirring 8 more hours.
The reaction mixture was filtered and the solvent removed in vacuo to give
title compound, 11.41 g (91%), as a white solid. MS (M+H).sup.+
=459.sup.+.
F.
N-1-(1,1-Dimethylethoxy)carbonyl!-4-piperidinyl!methyl!-1-N-(2,2,2-tr
ifluoro-ethyl)sulfonyl!-D-phenylalanyl!-L-prolinamide
To a solution of Part E compound (1.71 g, 3.5 mmol) and triethylamine (2.5
mL, 17.9 mmol) in chloroform (10 mL) at 0.degree. C. under argon was added
trifluoroethylsulfonyl chloride (0.5 mL, 4.5 mmol). The reaction mixture
was stirred to room temperature overnight. After 24 hr, triethylamine (2.0
mL, 14.3 mmol) and trifluoroethylsulfonyl chloride (0.5 mL, 4.5 mmol) were
added at 0.degree. C. The ice-water bath was removed, and the reaction was
stirred for 6 h. The mixture was diluted with 0.25M aqueous KHSO.sub.4 and
EtOAc. After separaton of the two layers, the organic layer was washed
with 0.25M aqueous KHSO.sub.4, H.sub.2 O, saturated aqueous NaHCO.sub.3,
and H.sub.2 O, successively; dried over NaSO.sub.4, filtered and
concentrated in vacuo to give title compound, (1.60 g, 75%): MS (M
+H).sup.+ =605.sup.+.
G.
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(2,2,2-trifluoroethyl
)sulfonyl!-D-phenylalanyl!-L-prolinamide, trifluoroacetate (1:1) salt
To a solution of Part F compound (1.60 g, 2.6 mmol) in dichloromethane (4.4
mL) at 0.degree. C. under argon was added trifluoroacetic acid (3.0 mL,
38.9 mmol). After 5.5 h, the reaction mixture was concentrated in vacuo to
give the corresponding TFA salt. To a solution of this salt (1.3 mmol) and
1H-pyrazole-1-carboxamidine hydrochloride (0.311 g, 2.1 mmol) in DMF (1.3
mL) under argon was added N,N-diisopropylethylamine (0.7 mL, 4.0 mmol).
After 2 days, 1H-pyrazole-1-carboxamidine hydrochloride (0.150 g, 1.0
mmol) and N,N-diisopropylethylamine (0.5 mL, 2.9 mmol) were added. After 1
day, 1H-pyrazole-1-carboxamidine hydrochloride (0.150 g, 1.0 mmol) and
N,N-diisopropylethylamine (0.5 mL, 2.9 mmol) were added. After 2 days, the
reaction mixture was concentrated in vacuo and purified by preparative
HPLC (CH.sub.3 CN/H.sub.2 O with TFA buffer using a C-18 silica gel
column). The appropriate fractions were combined, concentrated in vacuo,
dissolved in H.sub.2 O and lyophilized to yield title compound (0.256 g,
29%) as a white solid:
a!.sub.D =-56.0 (C 0.50, MeOH)
MS (M+H).sup.+ =-547.sup.+
Anal. Calc'd for C.sub.23 H.sub.33 N.sub.6 O.sub.4 SF.sub.3.1.22 TFA.1.14
H.sub.2 O: C, 43.26; H, 5.21; N, 11.90; S, 4.54; F, 17.92 Found: C, 42.82;
H, 4.87; N, 11.48; S, 4.98; F, 17.52
EXAMPLE 34
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(phenylmethyl)sulfonyl
!-L-alanyl!-L-prolinamide, trifluoroacetate (1:1) salt
A. N-(t-Butoxy)-carbonyl!-L-alanyl!-L-proline methyl ester
To a solution of N-BOC-L-alanine (150 mmol) in DMF (250 mL) at 0.degree.
C., is added proline methyl ester.HCl (150 mmol), 1-hydroxybenzotriazole
hydrate (22.2 g, 0.164 mmol), ethyl-3-(3-dimethylamino)propyl
carbodiimide.HCl (31.4 g, 0.164 mmol) and 4-methylmorpholine (22.6 g,
0.224 mmol). The reaction mixture is stirred for 12 h while allowing the
reaction to warm to room temperature. The reaction mixture is poured in
water (750 mL) and extracted with ethyl acetate (2.times.150 mL). The
combined organic extracts are washed as follows: KHSO.sub.4 (0.25M,
2.times.50 mL), water (1.times.50 mL), saturated aqueous NaHCO.sub.3
(2.times.50 mL) and saturated NaCl (1.times.50 mL). The organic layer is
dried over MgSO.sub.4, filtered and the solvent removed in vacuo to give
title compound.
B. N-(Phenylmethyl)sulfonyl!carbonyl!-L-alanyl!-L-proline methyl ester
A solution of Part A compound (5.2 mmol) in trifluoroacetic acid (3 mL) is
stirred at 0.degree. C. for 1.5 hr and and concentrated in vacuo to give
the corresponding TFA salt. This salt is dissolved in chloroform (10 mL),
and triethylamine (2.2 mL, 15.6 mmol) and benzylsulfonyl chloride (1.48,
7.8 mmol) at 0.degree. C. are added to it. The reaction mixture is stirred
to room temperature overnight. After 25 h, the reaction mixture is poured
into EtOAc (50 mL), washed with 0.25M aqueous KHSO.sub.4 (2.times.20 mL),
H.sub.2 O (1.times.20 mL) and saturated aqueous NaHCO.sub.3 (2.times.20
mL), successively; dried over MgSO.sub.4, filtered and concentrated in
vacuo to give title compound.
C. N-(phenylmethyl)sulfonyl!carbonyl!-L-alanyl!-L-proline
To a solution of Part B compound (3.75 mmole) in THF (10 mL) at 0.degree.
C. is added aqueous NaOH (1.0N, 9.5 mL). After 30 min the reaction mixture
is warmed to room temperature and stirring continued for an additional 4
h. The organic solvent is removed in vacuo, the aqueous layer acidified to
pH of 2-3 and extracted with ethyl acetate (3.times.10 mL). The combined
extracts are dried over MgSO.sub.4, filtered and the solvent removed in
vacuo to give title compound.
D.
N-1-(1,1-Dimethylethoxy)carbonyl!-4-piperidinyl!methyl!-1-N-(phenylme
thyl)sulfonyl!-L-alanyl!-L-prolinamide
To a solution of Part C compound (3.2 mmol) in DMF (15 mL) at 0.degree. C.
is added N1-BOC-4-methylamino piperidine (3.2 mmol),
1-hydroxybenzotriazole hydrate (0.476 g, 3.5 mmol),
ethyl-3-(3-dimethylamino)-propyl carbodiimide.HCl (0.675 g, 3.5 mmol) and
4-methylmorpholine until basic by pH paper. The reaction mixture is
stirred for 12 h while allowing the reaction to warm to room temperature.
The reaction mixture is poured into 0.25M aqueous KHSO.sub.4 (50 mL) and
extracted with EtOAc (2.times.20 mL). The organic layers are combined and
washed with 0.25M aqueous KHSO.sub.4 (1.times.30 mL), water (1.times.25
mL), saturated aqueous NaHCO.sub.3 (2.times.25 mL) and water (1.times.20
mL), the organic layer dried over MgSO.sub.4, filtered and the solvent
removed in vacuo to give title compound.
E.
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(phenylmethyl)sulfony
l!-L-alanyl!-L-prolinamide, trifluoroacetate (1:1) salt
A solution of Part D compound (2.2 mmol) in trifluoroacetic acid (10.0 mL)
is stirred at 0.degree. C. for 5 h. The reaction mixture is concentrated
in vacuo to give the corresponding TFA salt. To a solution of this salt
(2.2 mmol), 1H-pyrazole-1-carboxamidine hydrochloride (0.474 g, 3.3 mmol)
and N,N-diisopropylethylamine (1.1 mL, 6.5 mmol) in DMF (3.0 mL) are added
and stirred for 3 days. The reaction mixture is concentrated in vacuo and
purified by preparative HPLC (CH.sub.3 CN/H.sub.2 O with TFA buffer using
a C-18 silica gel column). The appropriate fractions are combined,
concentrated in vacuo, dissolved in H.sub.2 O and lyophilized to yield
title compound.
EXAMPLE 35
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(phenylmethyl)sulfonyl
!-glycyl!-L-prolinamide, trifluoroacetate (1:1) salt
A. N-(t-Butoxy)-carbonyl!-glycyl!-L-proline methyl ester
To a solution of N-BOC-glycine (150 mmol) in DMF (250 mL) at 0.degree. C.,
is added proline methyl ester.HCl (150 mmol), 1-hydroxybenzotriazole
hydrate (22.2 g, 0.164 mmol), ethyl-3-(3-dimethylamino)propyl
carbodiimide.HCl (31.4 g, 0.164 mmol) and 4-methylmorpholine (22.6 g,
0.224 mmol). The reaction mixture is stirred for 12 h while allowing the
reaction to warm to room temperature. The reaction mixture is poured in
water (750 mL) and extracted with ethyl acetate (2.times.150 mL). The
combined organic extracts are washed as follows: KHSO.sub.4 (0.25M,
2.times.50 mL), water (1.times.50 mL), saturated aqueous NaHCO.sub.3
(2.times.50 mL) and saturated NaCl (1.times.50 mL). The organic layer is
dried over MgSO.sub.4, filtered and the solvent removed in vacuo to give
title compound.
B. N-(Phenylmethyl)sulfonyl!carbonyl!glycyl!-L-proline methyl ester
A solution of Part A compound (5.2 mmol) in trifluoroacetic acid (3 mL) is
stirred at 0.degree. C. for 1.5 hr and and concentrated in vacuo to give
the corresponding TFA salt. This salt is dissolved in chloroform (10 mL),
and triethylamine (2.2 mL, 15.6 mmol) and benzylsulfonyl chloride (1.48,
7.8 mmol) at 0.degree. C. are added to it. The reaction mixture is stirred
to room temperature overnight. After 25 h, the reaction mixture is poured
into EtOAc (50 mL), washed with 0.25M aqueous KHSO.sub.4 (2.times.20 mL),
H.sub.2 O (1.times.20 mL) and saturated aqueous NaHCO.sub.3 (2.times.20
mL), successively; dried over MgSO.sub.4, filtered and concentrated in
vacuo to give title compound.
C. N-(Phenylmethyl)sulfonyl!carbonyl!glycyl!-L-proline
To a solution of Part B compound (3.75 mmole) in THF (10 mL) at 0.degree.
C. is added aqueous NaOH (1.0N, 9.5 mL). After 30 min the reaction mixture
is warmed to room temperature and stirring continued for an additional 4
h. The organic solvent is removed in vacuo, the aqueous layer acidified to
pH of 2-3 and extracted with ethyl acetate (3.times.10 mL). The combined
extracts are dried over MgSO.sub.4, filtered and the solvent removed in
vacuo to give title compound.
D.
N-1-(1,1-Dimethylethoxy)carbonyl!-4-piperidinyl!methyl!-1-N-(phenylme
thyl)sulfonyl!-glycyl!-L-prolinamide
To a solution of Part C compound (3.2 mmol) in DMF (15 mL) at 0.degree. C.
is added N1-BOC-4-methylamino piperidine (3.2 mmol),
1-hydroxybenzotriazole hydrate (0.476 g, 3.5 mmol),
ethyl-3-(3-dimethylamino)-propyl carbodiimide.HCl (0.675 g, 3.5 mmol) and
4-methylmorpholine until basic by pH paper. The reaction mixture is
stirred for 12 h while allowing the reaction to warm to room temperature.
The reaction mixture is poured into 0.25M aqueous KHSO.sub.4 (50 mL) and
extracted with EtOAc (2.times.20 mL). The organic layers are combined and
washed with 0.25M aqueous KHSO.sub.4 (1.times.30 mL), water (1.times.25
mL), saturated aqueous NaHCO.sub.3 (2.times.25 mL) and water (1.times.20
mL), the organic layer dried over MgSO.sub.4, filtered and the solvent
removed in vacuo to give title compound.
E.
N-1-(Aminoiminomethyl!-4-piperidinyl!methyl!-1-N-(phenylmethyl)sulfony
l!glycyl!-L-prolinamide, trifluoroacetate (1:1) salt
A solution of Part D compound (2.2 mmol) in trifluoroacetic acid (10.0 mL)
is stirred at 0.degree. C. for 5 h. The reaction mixture is concentrated
in vacuo to give the corresponding TFA salt. To a solution of this salt
(2.2 mmol), 1H-pyrazole-1-carboxamidine hydrochloride (0.474 g, 3.3 mmol)
and N,N-diisopropylethylamine (1.1 mL, 6.5 mmol) in DMF (3.0 mL) are added
and stirred for 3 days. The reaction mixture is concentrated in vacuo and
purified by preparative HPLC (CH.sub.3 CN/H.sub.2 O with TFA buffer using
a C-18 silica gel column). The appropriate fractions are combined,
concentrated in vacuo, dissolved in H.sub.2 O and lyophilized to yield
title compound.
Examples 36 to 53
Following the procedures of Examples 30 to 33, the following examples of
compounds of the invention may be prepared.
TABLE
__________________________________________________________________________
##STR80##
Example No.
R.sup.3 R R.sup.2
R.sup.1
n p o A R.sup.4
__________________________________________________________________________
36 CH.sub.3 CH.sub.2 OH(S)
H CH.sub.3
1 0 --
##STR81#
##STR82##
37 C.sub.6 H.sub.5
H OH H 0 1 CO
##STR83#
##STR84##
38
##STR85## CH.sub.2 C.sub.6 H.sub.5 (R)
OCH.sub.3
CH.sub.3
0 2 --
##STR86#
##STR87##
39
##STR88## CH.sub.2 C.sub.6 H.sub.5 (S)
40
##STR89## CH.sub.2 CH.sub.2 CONH.sub.2 (S)
H H 1 0 --
##STR90##
CH.sub.2 NH.sub.2
41 C.sub.6 H.sub.5
CH.sub.2 CH.sub.2 CONH.sub.2 (R)
H CH.sub.3
1 1 CO
##STR91##
CH.sub.2 NH.sub.2
42
##STR92## CH(OH)CH.sub.3 (S-Thr)
CHCH.sub.2 CH.sub.2 CH
0 2 --
##STR93#
##STR94##
43
##STR95## CH(OH)CH.sub.3 (S-alloThr)
CH.sub.3
H 1 2 CO
##STR96#
##STR97##
44
##STR98##
##STR99## SCH.sub.3
CH.sub.3
1 1 --
##STR100
#
##STR101##
45
##STR102##
CH.sub.2 CH.sub.2 CO.sub.2 H(R)
H CH.sub.3
0 0 CO
##STR103
#
##STR104##
46 C.sub.2 H.sub.5
CH.sub.2 OCH.sub.2 Ph(R)
H H 1 0 --
##STR105
#
##STR106##
47 C.sub.6 H.sub.5 CH.sub.2
CH.sub.2 CH.sub.2 Ph(S)
H H 1 1 --
##STR107
#
##STR108##
48
##STR109##
##STR110## H H 1 2 --
##STR111
#
##STR112##
__________________________________________________________________________
EXAMPLE 49
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(ethylsulfonyl)-D-pheny
lalanyl!-L-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =-49.2 (c 1.08, MeOH)
MS (M+H).sup.+ =493.sup.+
Anal. Calc'd for C.sub.23 H.sub.36 N.sub.6 O.sub.4 S.1.4 TFA.1.0 H.sub.2 O:
C, 46.23; H, 5.92; N, 12.54; S, 4.78; F, 11.90 Found: C, 46.23; H, 6.08;
N, 12.39; S, 4.71; F, 11.98
EXAMPLE 50
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(propylsulfonyl)-D-phen
ylalanyl!-L-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =-47.0 (c 1.02, MeOH)
MS (M+H).sup.+ =507.sup.+
Anal. Calc'd for C.sub.24 H.sub.38 N.sub.6 O.sub.4 S.1.15 TFA.1.2 H.sub.2
O: C, 47.91; H, 6.35; N, 12.74; S, 4.86; F, 9.94 Found: C, 47.90; H, 6.21;
N, 12.50; S, 4.82; F, 10.06
EXAMPLE 51
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(phenylmethyl)sulfonyl
!-D-phenylalanyl!-L-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =-44.6 (c 0.8, MeOH)
MS (M+H).sup.+ =555.sup.+
Anal. Calc'd for C.sub.28 H.sub.38 N.sub.6 O.sub.4 S.1.55 TFA.0.67 H.sub.2
O: C, 50.31; H, 5.42; N, 11.32; S, 4.32; F, 11.90 Found: C, 50.31; H,
5.69; N, 11.24; S, 4.26; F, 11.97
EXAMPLE 52
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(phenylsulfonyl)-D-phen
ylalanyl!-L-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =+10.1 (c 1.0, MeOH)
MS (M+H).sup.+ =541.sup.+
Anal. Calc'd for C.sub.27 H.sub.36 N.sub.6 O.sub.4 S.1.3 TFA.0.83 H.sub.2
O: C, 50,51; H, 5.58; N, 11.94; S, 4.55; F, 10.53 Found: C, 50.51; H,
5.47; N, 11.83; S, 4.71; F, 10.52
EXAMPLE 53
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(1-methylethyl)sulfony
l!-D-phenylalanyl!-L-prolinamide, trifluoroacetate (2:3) salt
a!.sub.D =-43.1 (c 1.10, MeOH)
MS (M+H).sup.+ =507.sup.+
Anal. Calc'd for C.sub.24 H.sub.38 N.sub.6 O.sub.4 S.1.5 TFA.1.35 H.sub.2
O: C, 46.20; H, 6.06; N, 11.97; S, 4.57; F, 12.18 Found: C, 46.61; H,
5.86; N, 11.95; S, 4.44; F, 12.02
EXAMPLE 54
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-((.+-.)-10-camphorsulfo
nyl)-D-phenylalanyl!-L-prolinamide, trifluoroacetate (1:1) salt
##STR113##
N-1-1,1,-Dimethylethoxy!carbonyl!amino1,1,-dimethylethoxy!carbonyl!i
mino!methyl!-4-piperidinyl!methyl!1-(N-carbobenzyloxy)-D-phenylalanyl!-L-p
rolinamide
To a stirred solution of
N-1-(1,1-dimethylethoxycarbonyl)-4-piperidinyl!methyl!-1-N-(carbobenzyl
oxy)-D-phenylalanyl!-L-prolinamide (10.0 g, 16.9 mmol) in 10 mL of dry
dichloromethane was added a solution of 4N HCl in dioxane (25 mL, 100
mmol). The reaction solution was stirred at room temperature for 2.5 h and
concentrated in vacuo to give the intermediate hydrochloride salt. To a
stirred solution of this salt, bis-t-butoxycarbonyl thiourea (5.12 g, 18.5
mmol) and triethyl amine (8.21 mL, 59.0 mmol) in 40 mL of DMF under argon
at 0.degree. C. was added HgCl.sub.2 (5.04 g, 18.5 mmol). The reaction
mixture was stirred at 0.degree. C. for 1 h and at room temperature for 30
min at which time another batch of triethyl amine (2.00 mL, 14.4 mmol),
bis-C-butoxycarbonyl thiourea (2.56 g, 9.25 mmol) and HgCl.sub.2 (2.52 g,
9.25 mmol) was added. The reaction mixture was stirred at room temperature
for 1 h and diluted with 0.5 L of EtOAc, filtered through a 3" pad of
Celite, the pad rinsed with EtOAc and the filtrate washed with 5%
KHSO.sub.4 solution and brine. The EtOAc layer was dried (MgSO.sub.4),
filtered and concentrated in vacuo to give the part A compound.
##STR114##
N-1-1,1,-Dimethylethoxy!carbonyl!amino1,1,-dimethylethoxy!carbonyl!i
minomethyl!-4-piperidinyl!methyl!-D-phenylalanyl-L-prolinamide
To a stirred solution of part A carbamate (3.60 g, 4.90 mmol) in 100 mL of
methanol under argon was added 20% Pd(OH).sub.2 /C (0.72 g). The
atmosphere was replaced with hydrogen, the mixture stirred at room
temperature for 20 h, the catalyst filtered off through a 4 mM
polycarbonate film, rinsed with methanol and concentrated in vacuo to give
2.41 g (82%) of part B amine.
##STR115##
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-((.+-.)-10-camphorsulfo
nyl)-D-phenylalanyl!-L-prolinamide, trifluoroacetate (1:1) salt
To a stirred solution of part B amine (600 mg, 1.00 mmol) and triethyl
amine (0.14 mL, 3.00 mmol) in 5 mL of chloroform under argon was added
(.+-.)-10-camphorsulfonyl chloride (376 mg, 1.50 mmol). The solution was
stirred at room temperature for 2 h at which time 0.20 mL (1.44 mmol) of
triethyl amine was added. The solution was stirred at room temperature for
25 h and diluted with EtOAc, washed with KHSO.sub.4 solution, saturated
NaHCO.sub.3 solution, and brine. The organic layer was dried (MgSO.sub.4),
filtered and concentrated in vacuo. To this bis-Bocguanidine (280 mg, 0.34
mmol) at 0.degree. C. was added TFA (3.00 mL, 38.9 mmol). The solution was
stirred at 0.degree. C. for 1 h and at room temperature for 1 h,
concentrated in vacuo and purified by preparative HPLC to give 240 mg
(93%) of
N-1-(aminoiminomethyl)-4-piperidinyl!methyl!-1-N-((.+-.)-10-camphorsulf
onyl)-D-phenylalanyl!-L-prolinamide, trifluoroacetate (1:1) salt.
.alpha.!.sub.D =-28.3.degree. (c=0.59, MeOH)
Anal. Calc'd for C.sub.31 H.sub.46 N.sub.6 O.sub.5 S.1.2 TFA: C, 53.37; H,
6.33; N, 11.18; S, 4.27; F, 9.10 Found: C, 53.16; H, 6.39; N, 10.93; S,
4.24; F, 8.98
EXAMPLE 55
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!1-N-(pentafluorophenylsulfon
yl)-D-phenylalanyl!-L-prolinamide, trifluoroacetate (1:1) salt
##STR116##
By substituting pentafluorophenyl sulfonyl chloride in the Example 1, Part
B reaction sequence there was obtained
N-1-(aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(pentafluorophenylsulf
onyl)-D-phenylalanyl!-L-prolinamide.
.alpha.!.sub.D =-18.4.degree. (c=0.63, MeOH)
Anal. Calc'd for C.sub.27 H.sub.31 N.sub.6 O.sub.4 SF.sub.5.1.00 TFA.1.00
H.sub.2 O: C, 45.67; H, 4.49; N, 11.02; S, 4.20; F, 19.93 Found: C, 45.71;
H, 4.24; N, 10.72; S, 4.38; F, 20.04
EXAMPLES 56 TO 63
Following the methods described above the following compounds were
prepared.
EXAMPLE 56
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!1-N-(pentamethylphenylsulfon
yl)-D-phenylalanyl!-L-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =+3.5.degree. (c=0.5, MeOH)
Anal. Calc'd for C.sub.32 H.sub.46 N.sub.6 O.sub.4 S.1.40 TFA.0.70 H.sub.2
O: C, 53.38 H, 6.28; N, 10.73; S, 4.09; F, 10.19 Found: C, 53.39; H, 6.00;
N, 10.41; S, 4.07; F, 9.93
EXAMPLE 57
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!1-N-(2,4,6-isopropylphenylsu
lfonyl)-D-phenylalanyl!-L-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =+21.7.degree. (c=0.5, MeOH)
Anal. Calc'd for C.sub.36 H.sub.54 N.sub.6 O.sub.4 S.1.40 TFA.0.40 H.sub.2
O: C, 55.89; H, 6.79; N, 10.08; S, 3.85; F, 9.57 Found: C, 55.91; H, 6.94;
N, 10.19; S, 3.97; F, 9.48
EXAMPLE 58
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!1-N-(4-carboxyphenylsulfonyl
)-D-phenylalanyl!-L-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =+21.5.degree. (c=0.79, MeOH)
Anal. Calc'd for C.sub.28 H.sub.36 N.sub.6 O.sub.4 S.1.00 TFA.0.73 H.sub.2
O: C, 50.62; H, 5.45; N, 11.80; S, 4.50; F, 8.01 Found: C, 50.83; H, 5.31;
N, 11.59; S, 4.66; F, 8.15
EXAMPLE 59
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!1-N-2,2,5,7,8-(pentamethylch
roman-6-sulfonyl)-D-phenylalanyl!-L-prolinamide, trifluoroacetate (1:1)
salt
.alpha.!.sub.D =+18.3.degree. (c=0.6, MeOH)
Anal. Calc'd for C.sub.35 H.sub.50 N.sub.6 O.sub.5 S.1.18 TFA.0.22 H.sub.2
O: C, 55.72; H, 6.46; N, 10.43; S, 3.98; F, 8.35 Found: C, 55.71; H, 6.57;
N, 10.27; S, 4.07; F, 8.36
EXAMPLE 60
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!1-N-(3-trifluoromethylphenyl
sulfonyl)-D-phenylalanyl!-L-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =+3.3.degree. (c=0.44, MeOH)
Anal. Calc'd for C.sub.28 H.sub.35 N.sub.6 O.sub.4 SF.sub.3.1.05 TFA.0.70
H.sub.2 O: C, 49.36; H, 5.02; N, 11.48; S, 4.38; F, 15.95 Found: C, 49.36;
H, 4.76; N, 11.24; S, 4.47; F, 15.98
EXAMPLE 61
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!1-N-(methylsulfonyl)-D-leucy
l!-L-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =-19.degree. (c=0.21, MeOH)
Anal. Calc'd for C.sub.19 H.sub.36 N.sub.6 O.sub.4 S.1.31 TFA.3.34 H.sub.2
O: C, 44.43; H, 5.44; N, 13.55; S, 5.17; F, 13.51 Found: C, 44.61; H,
5.28; N, 12.66; S, 4.98; F, 13.82
EXAMPLE 62
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(methylsulfonyl)-D-phen
ylalanyl!-L-pipecolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =-60.1.degree. (c=0.7, MeOH)
Anal. Calc'd for C.sub.23 H.sub.36 N.sub.6 O.sub.4 S.1.30 TFA.01.00 H.sub.2
O: C, 46.67 H, 6.01; N, 12.76; S, 4.87; F, 11.25 Found: C, 46.42; H, 5.58;
N, 12.52; S, 4.59; F, 11.02
EXAMPLE 63
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-N-(3-carboxyphenylsulfony
l)-D-phenylalanyl!-L-prolinamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =+8.3.degree. (c=0.84, MeOH)
Anal. Calc'd for C.sub.28 H.sub.36 N.sub.6 O.sub.6 S.1.03 TFA.0.71 H.sub.2
O: C, 50.50; H, 5.42; N, 11.76; S, 4.48; F, 8.21 Found: C, 50.50; H, 5.16;
N, 11.65; S, 4.60; F, 8.24
The following Examples represent preferred embodiments of the second
embodiment of the present invention. Unless otherwise indicated, all
temperatures are expressed in degrees Centigrade.
EXAMPLE I
S-(R*,R*)!-N-2-2-4-(Aminoiminomethyl)amino!butyl!thio!methyl!-1-pyrr
olidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
trifluoroacetate (1:1) salt
A.
(S)-2-4-(1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)butyl!thio!methyl!-1-py
rrolidinecarboxylic acid, 1,1-dimethylethyl ester
To a stirred solution of
(S)-2-(4-methylphenyl)sulfonyl!oxy!methyl!-1-pyrrolidinecarboxylic
acid, 1,1-dimethylethyl ester (reported in Journal of Med. Chemistry,
p2615, 1992) (10.0 g, 28.2 mmol) in 100 mL of acetone under argon was
added potassium thioacetate (3.91 g, 34.2 mmol). The reaction solution was
refluxed for 18 h at which time another batch of potassium thioacetate
(3.22 g, 28.2 mmol) was added. This mixture was refluxed for 24 h and
cooled to room temperature. The precipitate was filtered off through a 2"
pad of Celite and rinsed with acetone (2.times.30 mL). The filtrate was
concentrated in vacuo. The residue was dissolved in 300 mL of ether and
washed with water (2.times.50 mL) and brine (1.times.50 mL). The organic
layer was dried (Na.sub.2 SO.sub.4), filtered and concentrated in vacuo to
give 6.75 g of the intermediate thioacetate. To a stirred solution of this
thioacetate (3.27 g, 12.6 mmol) in 100 mL of methanol under argon at
0.degree. C. was added a solution of 1M t-C.sub.4 H.sub.9 OK in
tetrahydrofuran (THF) (12.6 mL, 12.6 mmol). This reaction solution was
then sparged with argon for 15 min. To this solution was added
N-(4-bromobutyl)phthalimide (3.56 g, 12.6 mmol). The reaction solution was
stirred at room temperature for 3 h and quenched at 0.degree. C. by
dropwise addition of 20 mL of saturated NH.sub.4 Cl solution. The mixture
was concentrated in vacuo. The residue was diluted with 600 mL of EtOAc
and washed with saturated NaHCO.sub.3 solution (2.times.200 mL) and brine
(1.times.200 mL). The EtOAc layer was dried (MgSO.sub.4), filtered and
concentrated in vacuo. Purification was effected by flash chromatography
on 180 g of Merck silica gel 60 using 29% ether in hexane as eluant to
give 3.16 g (55%) of title thioether.
B.
S-(R*,R*)!-2-2-4-(1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)butyl!thio!
methyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!carbamic acid,
1,1-dimethylethyl ester
To a stirred solution of Part A thioether (3.28 g, 7.85 mmol) in 10 mL of
dry dichloromethane was added 10 mL of trifluoroacetic acid (TFA). The
solution was stirred at room temperature for 1.5 h and diluted with 100 mL
of ether. The solution was concentrated in vacuo to give a crude TFA.amine
salt. To a stirred solution of this salt, 1-hydroxybenzotriazole
monohydrate (HOBT) (1.33 g, 7.85 mmol) and N-Boc-L-serine (1.70 g, 7.85
mmol) in 40 mL of dimethylformamide (DMF) was added in order
N-methylmorpholine (NMM) (2.58 mL, 23.5 mmol) and
ethyl-3-(3-dimethylamino)propyl carbodiimide hydrochloride (1.50 g, 7.85
mmol). This reaction solution was sparged with argon for 10 min and
stirred at room temperature for 18 h. The mixture was concentrated under
pump vacuum at 45.degree. C. The residue was dissolved in 200 mL of EtOAc
and washed with 1N HCl solution (2.times.80 mL), saturated NaHCO.sub.3
solution (1.times.80 mL) and brine (1.times.80 mL). The EtOAc layer was
dried (MgSO.sub.4), filtered and concentrated in vacuo to give 3.65 g
(90%) of title carbamate.
C.
S-(R*,R*)!-N-2-2-4-(1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)butyl!thi
o!methyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfon
amide
To a stirred solution of Part B carbamate (3.65 g, 7.06 mmol) in 15 mL of
dry dichloromethane was added 15 mL of TFA. The solution was stirred at
room temperature for 1.5 h and diluted with 200 mL of ether. The solution
was concentrated in vacuo to give a crude TFA,amine salt. To a stirred
solution of this salt and triethyl amine (2.95 mL, 21.2 mmol) in 60 mL of
dry dichloromethane at 0.degree. C. was added 2-naphthalenesulfonyl
chloride (1.76 g, 7.77 mmol). The reaction solution was stirred at room
temperature for 4.5 h and diluted with 140 mL of dichloromethane. The
solution was washed with 1N HCl solution (2.times.60 mL), saturated
NaHCO.sub.3 solution (2.times.60 mL) and brine (1.times.60 mL). The
organic layer was dried (MgSO.sub.4), filtered and concentrated in vacuo.
This was chromatographed on 120 g of Merck silica gel 60 using 2% CH.sub.3
OH/CH.sub.2 Cl.sub.2 as eluant to give 2.62 g (61%) of title sulfonamide.
D.
S-(R*,R*)!-N-2-2-(4-Aminobutyl)thio!methyl!-1-pyrrolidinyl!-1-(hydrox
ymethyl)-2-oxoethyl!-2-naphthalenesulfonamide
To a stirred solution of Part C sulfonamide (0.77 g, 1.27 mmol) in 20 mL of
dry dichloromethane under argon was added anhydrous hydrazine (0.24 g,
7.61 mmol). The reaction mixture was stirred at room temperature for 2.5 h
and concentrated in vacuo. The residue was combined with 30 mL of methanol
and 60 mL of toluene. The mixture was concentrated in vacuo. This
co-evaporation with methanol and toluene was repeated three times. The
residue was dissolved in 80 mL of methanol and the solution was heated to
reflux for 17 h. The mixture was then cooled to room temperature and
acidified to pH 1 by the addition of 1N HCl in ether. The mixture was
cooled in an acetone dry ice bath for 5 min and the precipitate was
filtered off, the solid was rinsed with methanol (2.times.10 mL), and the
filtrate was concentrated in vacuo. The product was purified by prep HPLC
and lyophilized to give 290 mg (39%) of title amine.TFA.
E.
S-(R*,R*)!-N-2-2-4-(Aminoiminomethyl)amino!butyl!thio!methyl!-1-pyr
rolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
trifluoroacetate (1:1) salt
To a stirred mixture of Part D amine.TFA (280 mg, 0.47 mmol) and Et.sub.3 N
(0.36 mL, 2.61 mmol) in 4 mL of absolute EtOH under argon was added
aminoiminomethanesulfonic acid (206 mg, 1.66 mmol). The mixture was
stirred at room temperature for 3 h and concentrated in vacuo. This was
purified by prep HPLC to give 180 mg (57%) of title compound.
Opt. Rotation: .alpha.!.sub.D =-54.7.degree. (c=0.70, MeOH).
Analysis calc'd for 0.33 H.sub.2 O+1.35 TFA: C, 46.24; H, 5.29; N, 10.49;
S, 9.61; F, 11.53 Found: C, 46.24; H, 5.22; N, 10.47; S, 9.92; F, 11.47.
EXAMPLE II
S-(R*,R*)!-N-4-1-(2-Amino-1-oxo-3-phenylpropyl)-2-pyrrolidinyl!methyl!
thio!butyl!guanidine, trifluoroacetate (2:5) salt
A.
S-(R*,S*)!-2-2-4-(1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)butyl!thio!
methyl!-1-pyrrolidinyl!-2-oxo-1-(phenylmethyl)ethyl!carbamic acid,
1,1-dimethylethyl ester
To a stirred solution of TFA.amine (prepared in Example I, Part B) (3.11g,
7.20 mmol), 1-hydroxybenzotriazole monohydrate (1.22 g, 7.20 mmol) and
N-Boc-D-phenylalanine (1.91 g, 7.20 mmol) in 60 mL of DMF was added in
order 4-methylmorpholine (3.16 mL, 28.8 mmol) and
ethyl-3-(3-dimethylamino)propyl carbodiimide hydrochloride (1.38 g, 7.20
mmol). This reaction solution was sparged with argon for 20 min and
stirred at room temperature for 18 h. The mixture was concentrated under
pump vacuum at 45.degree. C. The residue was dissolved in 400 mL of EtOAc
and washed with 1N HCl solution (2.times.100 mL), saturated NaHCO.sub.3
solution (2.times.100 mL) and brine (1.times.100 mL). The EtOAc layer was
dried (MgSO.sub.4), filtered and concentrated in vacuo. Purification was
effected by flash chromatography to give 2.74 g (68%) of title carbamate.
B.
S-(R*,S*)!-2-2-(4-Aminobutyl)thio!methyl!-1-pyrrolidinyl!-2-oxo-1-(ph
enylmethyl)ethyl!carbamic acid, 1,1-dimethylethyl ester
To a stirred solution of Part A carbamate (1.01 g, 1.79 mmol) in 25 mL of
methanol under argon was added anhydrous hydrazine (1.00 mL, 31.9 mmol).
The reaction solution was heated at 50.degree. C. for 3 h and concentrated
in vacuo. The residue was combined with 50 mL of methanol and 20 mL of
toluene. The mixture was concentrated in vacuo. The residue was diluted
with 40 mL of 1N NaOH solution and extracted with EtOAc (3.times.70 mL).
The combined organic extracts were dried (MgSO.sub.4), filtered and
concentrated in vacuo to give 700 mg (89%) of title amine.
C.
S-(R*,S*)!-2-2-4-(Aminoiminomethyl)-amino!butyl!thio!methyl!-1-pyrr
olidinyl!-2-oxo-1-(phenylmethyl)ethyl!carbamic acid, 1,1-dimethylethyl
ester
To a stirred mixture of Part B amine (680 mg, 1.56 mmol) and triethylamine
(Et.sub.3 N) (0.76 mL, 5.47 mmol) in 7 mL of absolute EtOH under argon was
added aminoimino-methanesulfonic acid (485 mg, 3.91 mmol). The mixture was
stirred at room temperature for 2 h concentrated in vacuo, and purified by
prep HPLC to give 780 mg (92%) of title BOC-amine.
D.
S-(R*,R*)!-N-4-1-(2-Amino-1-oxo-3-phenylpropyl)-2-pyrrolidinyl!methyl
!thio!butyl!guanidine, trifluoroacetate (2:5) salt
To Part C BOC.amine (410 mg, 0.69 mmol) at 0.degree. C. was added 4 mL of
TFA. The reaction solution was stirred at room temperature for 30 min and
diluted with 20 mL of methanol. The solution was concentrated in vacuo and
purified by prep HPLC to give 130 mg (28%) of title aminoguanidine.
Opt. rotation: .alpha.!.sub.D =-73.3.degree. (c=0.54, MeOH).
Analysis calc'd for 1.03 H.sub.2 O+2.30 TFA: C, 43.06; H, 5.41; N, 10.64;
S, 4.87; F, 19.91 Found: C, 43.06; H, 5.14; N, 10.40; S, 5.22; F, 20.00.
EXAMPLE II
S-(R*,R*)!-N-2-2-4-(Aminoiminomethyl)amino!butyl!sulfonyl!methyl!-1-p
yrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
rifluoroacetate (1:1) salt
A.
S-(R*,R*)!-N-2-2-4-(1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)butyl!sul
fonyl!methyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesu
lfonamide
To a stirred solution of the thioether prepared in Example I, Part C (1.00
g, 1.65 mmol) in 50 mL of dichloromethane was added m-chloroperbenzoic
acid (MCPBA) (1.14 g, 3.29 mmol). The reaction solution was stirred at
room temperature for 10 min and quenched by the addition of 10% sodium
bisulfite solution until the KI-starch paper test was negative. The
mixture was diluted with 300 mL of EtOAc and washed with saturated
NaHCO.sub.3 solution (2.times.150 mL) and brine (1.times.150 mL). The
organic layer was dried (MgSO.sub.4), filtered and concentrated in vacuo
to give 1.20 g (97%) of title sulfone.
B.
S-(R*,R*)!-N-2,2-(4-Aminobutyl)sulfonyl!methyl!-1-pyrrolidinyl!-1-(hy
droxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide
To a stirred solution of Part A sulfone (1.20 g, 1.96 mmol) in 30 mL of
methanol under argon was added anhydrous hydrazine (0.63 g, 19.6 mmol).
The reaction solution was heated to reflux for 2.5 h and concentrated in
vacuo. The residue was combined with 40 mL of methanol and 80 mL of
toluene. The mixture was concentrated in vacuo, the residue was diluted
with 40 mL of 1N NaOH solution and extracted with EtOAc (2.times.80 mL)
and 10% isopropyl alcohol (IPA) in dichloromethane (3.times.80 mL). The
combined organic extracts were dried (MgSO.sub.4), filtered, concentrated
in vacuo and purified by prep HPLC to give 350 mg (30%) of title
amine.TFA.
C.
S-(R*,R*)!-N-2-2-4-(Aminoiminomethyl)amino!butyl!sulfonyl!methyl!-1
-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
trifluoroacetate (1:1) salt
To a stirred mixture of Part B amine.TFA (340 mg, 0.57 mmol) and Et.sub.3 N
(0.32 mL, 2.29 mmol) in 6 mL 20 of absolute EtOH under argon was added
aminoiminomethanesulfonic acid (177 mg, 1.43 mmol). The mixture was
stirred at room temperature for 4 h concentrated in vacuo, and purified by
prep HPLC to give 370 mg (92%) of title compound.
Opt. rotation: .alpha.!.sub.D =-53.3.degree. (c=0.95, MeOH).
Analysis calc'd for 0.16 H.sub.2 O+1.40 TFA: C, 44.13; H, 4.98; N, 9.97; S,
9.13; F, 11.36 Found: C, 44.13; H, 4.87; N, 9.97; S, 9.05; F, 11.48.
EXAMPLE IV
S-(R*,R*)!-N-2-2-3-(Aminoiminomethyl)amino!propyl!thio!methyl!-1-pyr
rolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
trifluoroacetate (1:1) salt
A.
(S)-2-3-(1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)propyl!thio!methyl!-1-p
yrrolidinecarboxylic acid, 1,1-dimethylethyl ester
To a stirred solution of the tosylate employed as a starting material in
Example I, Part A (10.0 g, 28.2 mmol) in 120 mL of acetone under argon was
added potassium thioacetate (4.14 g, 36.2 mmol). The reaction solution was
sparged with argon for 10 min. This mixture was refluxed for 4 h and
cooled to room temperature. The precipitate was filtered off through a 3"
pad of Celite and rinsed with acetone (3.times.30 mL). The filtrate was
concentrated in vacuo. The residue was dissolved in 400 mL of ether and
washed with cold water (1.times.100 mL) and brine (1.times.100 mL). The
organic layer was dried (Na.sub.2.sub.SO.sub.4), filtered and concentrated
in vacuo to give 7.20 g (99%) of the intermediate thioacetate. To a
stirred solution of this thioacetate (7.20 g, 27.8 mmol) in 250 mL of
methanol under argon at 0.degree. C. was added a solution of 1M t-BuOK in
THF (27.8 mL, 27.8 mmol). This reaction solution was then sparged with
argon for 15 min. To this solution was added N-(3-bromopropyl)-phthalimide
(7.45 g, 27.8 mmol). The reaction solution was stirred at room temperature
for 4 h and quenched at 0.degree. C. by dropwise addition of 40 mL of
saturated NH.sub.4 Cl solution. The mixture was concentrated in vacuo, the
residue diluted with 400 mL of EtOAc and washed with saturated NaHCO.sub.3
solution (2.times.100 mL) and brine (1.times.100 mL). The EtOAc layer was
dried (MgSO.sub.4), filtered and concentrated in vacuo. Purification was
effected by flash chromatography on to give 6.11 g (54%) of title
thioether.
B.
S-(R*,R*)!-2-2-3-(1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)propyl!thio
!methyl!-1-pyrrolidinyl-1-(hydroxymethyl)-2-oxoethyl!carbamic acid,
1,1-dimethylethyl ester
To a stirred solution of Part A thioether (6.00 g, 14.9 mmol) in 10 mL of
dry dichloromethane was added 15 mL of TFA. The solution was stirred at
room temperature for 1.5 h and diluted with 150 mL of ether. The solution
was concentrated in vacuo to give a crude TFA.amine salt. To a stirred
mixture of this TFA.amine salt in 100 mL of dry dichloromethane was added
a solution of 1N HCl in ether (20.0 mL, 20.0 mmol). The mixture was
concentrated in vacuo to give the amine.HCl salt in a quantitative yield.
To a stirred solution of this HCl salt (4.05 g, 9.19 mmol),
1-hydroxybenzotriazole monohydrate (1.55 g, 9.19 mmol) and N-Boc-L-serine
(2.00 g, 9.19 mmol) in 40 mL of DMF was added in order 4-methylmorpholine
(4.04 mL, 36.8 mmol) and ethyl-3-(3-dimethylamino)propyl carbodiimide
hydrochloride (1.76 g, 9.19 mmol). This reaction solution was sparged with
argon for 10 min and stirred at room temperature for 22 h. The mixture was
concentrated under pump vacuum at 45.degree. C. The residue was dissolved
in 300 mL of EtOAc and washed with 1N HCl solution (2.times.100 mL),
saturated NaHCO.sub.3 solution (1.times.100 mL) and brine (1.times.100
mL). The EtOAc layer was dried (MgSO.sub.4), filtered and concentrated in
vacuo to give 3.88 g (84%) of title carbamate.
C.
S-(R*,R*)!-N-2-2-3-(1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)propyl!th
io!methyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfo
namide
To a stirred solution of Part B carbamate (2.04 g, 4.06 mmol) in 10 mL of
dry dichloromethane was added at 0.degree. C. a solution of 4N HCl in
dioxane (15.0 mL, 60.0 mmol). The solution was stirred at room temperature
for 1.5 h and diluted with 200 mL of ether. The solution was concentrated
in vacuo to give a crude HCl.amine salt. To a stirred solution of this
salt and triethyl amine (1.69 mL, 12.2 mmol) in 60 mL of dry
dichloromethane at 0.degree. C. was added 2-naphthalenesulfonyl chloride
(1.01 g, 4.46 mmol). The reaction solution was sparged with argon for 5
min and stirred at room temperature for 3 h. The reaction solution was
diluted with 240 mL of dichloromethane. The solution was washed with 1N
HCl solution (3.times.60 mL), saturated NaHCO.sub.3 solution (2.times.60
mL) and brine (1.times.100 mL). The organic layer was dried (MgSO.sub.4),
filtered and concentrated in vacuo. The residue was washed with hexane
(2.times.60 mL) and the solid was chromatographed on 120 g of Merck silica
gel 60 using 3% CH.sub.3 OH/CH.sub.2 Cl.sub.2 as eluant to give 1.69 g
(70%) of title sulfonamide.
D.
S-(R*,R*)!-N-2-2-3-(Aminoiminomethyl)amino!propyl!thio!methyl!-1-py
rrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
trifluoroacetate (1:1) salt
To a stirred solution of Part C sulfonamide (1.59 g, 1.68 mmol) in 40 mL of
methanol under argon was added anhydrous hydrazine (0.54 g, 16.8 mmol).
The reaction mixture was heated at 45.degree. C. for 5 h and concentrated
in vacuo. The residue was combined with 60 mL of methanol and 100 mL of
toluene. The mixture was concentrated in vacuo to give 1.07 g (85%) of the
intermediate amine. To a stirred mixture of this amine (1.05 g, 2.25 mmol)
and Et.sub.3 N (1.09 mL, 7.87 mmol) in 50 mL of absolute EtOH under argon
was added aminoiminomethanesulfonic acid (698 mg, 5.62 mmol). The mixture
was stirred at room temperature for 4 h, concentrated in vacuo and
purified by prep HPLC to give 350 mg (25%) of title compound.
Opt. rotation: .alpha.!.sub.D =-63.1.degree. (c=1.12, MeOH).
Analysis calc'd for 0.30 H.sub.2 O+1.07 TFA: C, 46.69; H, 5.30; N, 11.28;
S, 10.32; F, 9.82 Found: C, 46.64; H, 5.24; N, 11.12; S, 10.37; F, 9.85.
EXAMPLE V
S-(R*,R*)!-N-2-2-1-(Aminoiminomethyl)-4-piperidinyl!methyl!thio!meth
yl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
trifluoroacetate (1:1) salt
A.
(S)-4-1-(1,1-Dimethylethoxy)carbonyl!-2-pyrrolidinyl!methyl!thio!meth
yl!-1-piperidinecarboxylic acid, phenylmethyl ester
To a solution of the thioacetate intermediate prepared in Example I, Part
A,
##STR117##
and 4-tosyloxymethyl-N-carbobenzyloxy piperidine, 1.5 g, 3.86 mmol) in 35
mL DMF was added NaOMe (0.88 mL, 15.44 mmol) at 0.degree. C. After the
reaction stirred for 10 minutes and the ice bath was removed, the reaction
was stirred at room temperature for 2 hrs. NaOMe (0.6 mL) was added and
the reaction stirred for another 30 minutes. The reaction mixture was
concentrated and partitioned between EtOAc and H.sub.2 O, the EtOAc layer
washed with brine, dried over Na.sub.2 SO.sub.4 and concentrated in vacuo
to give the crude product which then was purified by a silica gel column
(EtOAc:hexane=1:4) (87% yield).
B. N-(2-Naphthalenylsulfonyl)-L-serine
To a solution of 2-naphthalene sulfonyl chloride (3 g, 13.23 mmol) and
serine benzylester (3.07 g, 13.23 mmol) in 120 mL CH.sub.2 Cl.sub.2 was
added Et.sub.3 N (5.53 mL, 13.23 mmol) at 0.degree. C. The reaction was
stirred at room temperature for 1 hr and washed with a saturated (sat.)
solution of KHSO.sub.4, brine, and dried over Na.sub.2 SO.sub.4. The
organic layer was concentrated in vacuo to give the naphthylsulfonamide
(93%).
To a solution of the naphthylsulfonamide (1 g, 2.59 mmol) and Pd/C (10%, 50
mg) in 25 mL EtOAc was added 2.6 mL of 1N HCl, followed by H.sub.2. The
reaction was stirred at room temperature for 16 hrs and the catalyst was
removed by filtration. The filtrate was concentrated in vacuo to give (89%
yield).
C.
S-(R*,R*)!-N-1-(Hydroxymethyl)-2-oxo-2-1-(phenylmethoxy)carbonyl!-
4-piperidinyl!methyl!thio!methyl!-1-pyrrolidinyl!ethyl!-2-naphthalenesulfon
amide
To a solution of Part A compound (1.39 g, 3.10 mmol) in 2 mL CH.sub.2
Cl.sub.2 was added 9.3 mL trifluoroacetic acid (TFA) at 0.degree. C. The
ice bath was removed and the reaction was stirred at room temperature for
30 minutes. The reaction mixture was concentrated in vacuo and
co-evaporated with Et.sub.2 O to give the TFA salt (70% yield).
To a solution of Part B compound (0.638 g, 2.16 mmol) and HOBT (0.321 g,
2.37 mmol) in 10 mL DMF was added WSC (0.46 g, 2.37 mmol) and the TFA salt
(100 g, 2.16 mmol), followed by NMM (0.264 mL, 2.37 mmol). The reaction
was stirred at room temperature for 16 hrs and partitioned between EtOAc
and a sat. solution of NaHCO.sub.3. The organic layer was further washed
with a sat. solution of KHSO.sub.4, brine, dried over Na.sub.2 SO.sub.4
and concentrated in vacuo to give (81% yield).
D.
S-(R*,R*)!-N-1-(Acetyloxy)methyl!-2-oxo-2-2-(4-piperidinylmethyl)thi
o!methyl!-1-pyrrolidinyl!ethyl!-2-naphthalenesulfonamide
To a solution of Part C compound (1.1 g, 2.08 mmol) in 1 mL CH.sub.2
Cl.sub.2 was added 6.24 mL HBr in HOAc (30%) at 0.degree. C. The ice bath
was removed and stirred at room temperature for 1 hr. The reaction mixture
was concentrated in vacuo and triturated with Et.sub.2 O to give title
compound.
E.
S-(R*,R*)!-N-1-(Acetyloxy)methyl!-2-2-1-(aminoiminomethyl)-4-piper
idinyl!methyl!thio!methyl!-1-pyrrolidinyl!-2-oxoethyl!-2-naphthalenesulfona
mide
To a solution of Part D (1.47 g, 2.57 mmol) and H-pyrazole-1-carboxamidine
(0.42 g, 2.83 mmol) in 8 mL DMF was added diisopropylethylamine (0.898 mL,
5.14 mmol). The reaction was stirred at room temperature for 16 hrs and
purified by a preparative HPLC to give title compound (25% yield).
F.
S-(R*,R*)!-N-2-2-1-(Aminoiminomethyl)-4-piperidinyl!methyl!thio!met
hyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide
, trifluoroacetate (1:1) salt
To a solution of Part E compound (0.33 g, 0.57 mmol) in 2 mL MeOH was added
1N NaOH (1.72 mL, 1.72 mmol) at 0.degree. C. The ice bath was removed and
stirred at room temperature for 1.5 hrs. The reaction mixture was
concentrated in vacuo and purified on a preparative HPLC to give title
compound (56% yield).
Anal. Calc'd for C.sub.25 H.sub.35 N.sub.5 O.sub.4 S.sub.2.1.2 TFA.1.2
H.sub.2 O: C, 47.55; H, 5.62; N, 10.12; S, 9.26 Found: C, 47.60; H, 5.36;
N, 9.99; S, 10.08.
.alpha.!.sub.D =-164.9 (c=3.04, CD.sub.3 OD).
EXAMPLE VI
S-(R*,R*)!-N-2-2-1-(Aminoiminomethyl)-4-piperidinyl!methoxy!methyl!-1-
pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide
A.
(S)-4-1-(1,1-Dimethylethoxy)carbonyl!-2-pyrrolidinyl!methoxy!methyl!-1
-piperidinecarboxylic acid, phenylmethyl ester
To BOC-L-prolinol (10 mmol) in xylene (10 mL) was added KOH (20 mmol),
4-tosyloxymethyl-N-carbobenzyloxy piperidine (11 mmol) and the reaction
was heated to reflux for 3 hours. The reaction mixture was stirred at room
temperature for 16 hours, quenched with NH.sub.4 Cl, extracted with ethyl
acetate and purified by column chromatography to provide the title ether.
B.
S-(R*,R*)!-4-1-3-Hydroxy-2-(2-naphthalenylsulfonyl)amino!-1-oxopropy
l!-2-pyrrolidinyl!methoxy!methyl!-1-piperidinecarboxylic acid, phenylmethyl
ester
The Part A ether (9 mmol) was dissolved in trifluoroacetic acid (10 mL) at
0.degree. C. After stirring for 30 min, the TFA was evaporated in vacuo
and the trifluoracetate salt precipitated with ether. The TFA salt was
dissolved in DMF (10 mL) and reacted with N-2-napthylsulfonyl-serine (9
mmol) in the presence of WSC (9 mmol), HOBT (9 mmol) and NMM (9 mmol).
After stirring for 12 hours, the reaction mixture was diluted with ethyl
acetate (50 mL), washed with sat'd KHSO.sub.4 (3.times.), sat'd
NaHCO.sub.3, sat'd NaCl, and dried in vacuo to provide the title
naphthylsulfonyl serine derivative.
C.
S-(R*,R*)!-N-2-2-1-(Aminoiminomethyl)-4-piperidinyl!methoxy!methyl!-
1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide
The Part B compound (8 mmol) was dissolved in HBr/HOAc at 0.degree. C. for
30 min. The reaction mixture was then made basic with NaOH and extracted
with ethyl acetate. The free base derived from the ethyl acetate layer was
treated with 1H-pyrazolecarboxamidine in ethanol at 40.degree. C. for 3
hours, the reaction mixture stripped and purified by preparative HPLC to
provide the title compound.
EXAMPLE VII
S-(R*,S*)!-N-2-2-4-(Aminoiminomethyl)amino!butyl!thio!methyl!-1-pyrr
olidinyl!-2-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide
The title compound was prepared employing a procedure similar to that
described in Example I except substituting
(R)-2-(4-methylphenyl)sulfonyl!oxy!methyl!-1-pyrrolidine carboxylic
acid, 1,1-dimethylethyl ester for the corresponding (S)-isomer (140 mg,
40% yield).
Anal. Calc'd for C.sub.23 H.sub.33 N.sub.5 O.sub.4 S.sub.2.1.1 TFA.0.04
H.sub.2 O: C, 47.81, H, 5.43; N, 11.06; S, 10.13; F, 9.90 Found: C, 47.75;
H, 5.35; N, 10.93; S, 10.14; F, 9.78
EXAMPLE VIII
S-R*,R*-(E)!!-N-2-2-(4-Amino-2-butenyl)thio!methyl!-1-pyrrolidinyl!-1-
(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide
A.
(S,Z)-2-4-(2-Pyrrolidinylmethyl)thio!-2-butenyl!-1H-isoindole-1,3(2H)-di
one, hydrochloride
To a solution of
(S,Z)-2-4-1-(1,1-dimethylethoxy)carbonyl!-2-pyrrolidinyl!methyl!thio!
2-butenyl!-1H-isoindole-1,3(2H)-dione, (prepared as in Example I, Part A,
except substituting 4-bromo-2-butenyl phthalimide for the corresponding
phthalimide) (3.60 mmol)) in 2 mL of CH.sub.2 Cl.sub.2 was added 18 mL of
4N HCl/dioxane at 0.degree. C. The ice bath was removed and the reaction
was stirred at room temperature for 3 hrs. The reaction mixture was
concentrated in vacuo to give title compound (92% yield).
B.
S-R*,R*-(Z)!!-N-2-2-4-(1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)-2-bu
tenyl!thio!methyl!-1-pyrrolidinyl!-2-(hydroxymethyl)-2-oxoethyl!-2-naphthal
enesulfonamide
To a solution of Example V, Part B compound (650 mg, 2.20 mmol) and HOBT
(327 mg, 2.42 mmol) in 14 mL of DMF was added WSC (464 mg, 2.42 mmol),
followed by the Part A amine salt (818 mg, 2.20 mmol). The pH was adjusted
to 8 by NMM (0.27 mL, 2.42 mmol) and the reaction was stirred at room
temperature for 16 hrs. The reaction mixture was partitioned between
H.sub.2 O and EtOAc and EtOAc was washed with a sat. solution of
NaHCO.sub.3, a sat. solution of KHSO.sub.4, brine and dried over
MgSO.sub.4. The organic layer was concentrated in vacuo to give title
compound (86% yield).
C.
S-R*,R*-(E)!!-N-2-2-(4-Amino-2-butenyl)thio!-methyl!-1-pyrrolidinyl!
-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide
To a solution of Part B compound (800 mg, 1.34 mmol) in 34 mL CH.sub.3 OH
was added hydrazine (0.42 mL, 13.4 mmol) at room temperature and the
reaction was stirred at 40.degree.-50.degree. C. for 3 hrs. The reaction
mixture was concentrated in vacuo and partitioned between EtOAc and 1N
NaOH; the aqueous layer was salted out with NaCl and extracted with EtOAc.
The organic layers were combined and concentrated in vacuo to give the
crude product which was crystallized in EtOAc (71% yield).
Anal. Calc'd for C.sub.22 H.sub.29 N.sub.3 O.sub.4 S.sub.2.1.00 HCl.0.14
H.sub.2 O: C, 52.57; H, 6.07; N, 8.36; S, 12.76; Cl, 7.05 Found: C, 52.84;
H, 6.01; N, 8.09; S, 12.57; Cl, 7.14
.alpha.!.sub.D =-36.8 (c=0.50, MeOH).
EXAMPLE IX
S-R*,R*-(E)!!-N-2-4-(Aminoiminomethyl)amino!-2-butenyl!thio!methyl!
-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
trifluoroacetate (1:) salt
To a solution of Example VIII title compound (0.4 g, 0.86 mmol) and
amidinesulfonic acid (161 mg, 1.29 mmol) in 5 mL EtOH was added Et.sub.3 N
(0.364 mL, 2.59 mmol). The reaction was stirred at room temperature for 1
hr. The reaction mixture was concentrated in vacuo and purified by
preparative HPLC to give title compound (60% yield).
Anal. Calc'd for C.sub.23 H.sub.31 N.sub.5 O.sub.4 S.sub.2.1.20 TFA.0.28
H.sub.2 O: C, 47.11; H, 5.10; N, 10.82; S, 9.90; F, 10.56 Found: C, 47.12;
H, 4.92; N, 10.56; S, 9.48; F, 10.23
.alpha.!.sub.D =-26.6 (c=0.50, MeOH).
EXAMPLE X
S-(R*,R*)!-N-2-2-1-(Aminoiminomethyl)-3-piperidinyl!methyl!thio!meth
yl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
tifluoroacetate (1:1) salt
The title compound was prepared in a manner similar to that described in
Example V, except substituting 3-tosyloxymethyl-N-carbobenzyloxy
piperidine for 4-tosyloxymethyl-N-carbobenzyloxy piperidine, to provide
the desired product.
Anal. Calc'd for C.sub.25 H.sub.35 N.sub.5 O.sub.4 S.sub.2.1.2 TFA.1.01
H.sub.2 O: C, 47.78; H, 5.59; N, 10.17; S, 9.31 Found: C, 47.78; H, 5.46;
N, 9.97; S, 9.37
.alpha.!.sub.D =-57.7 (c=2.01, DMSO).
EXAMPLE XI
S-(R*,R*)!-N-2-2-5-(Aminoiminomethyl)amino!pentyl!thio!methyl!-1-pyr
rolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-1-naphthalenesulfonamide,
trifluoroacetate (1:1) salt
The title compound was prepared as described in Example I, except
N-(5-bromopentyl)phthalimide was substituted for the
N-(4-bromobutyl)phthalimide.
Anal. Calc'd for C.sub.24 H.sub.35 N.sub.5 O.sub.4 S.1.16 TFA.0.92 H.sub.2
O: C, 47.15; H, 5.71; N, 10.44; S, 9.56 Found: C, 47.15; H, 5.42; N,
10.36; S, 9.91
.alpha.!.sub.D =-263.7 (c=4.51, CD.sub.3 OD).
EXAMPLE XII
S-(R*,R*)!-N-2-2-6-(Aminoiminomethyl)amino!hexyl!thio!methyl!-1-pyr
rolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
trifluoroacetate (1:1) salt
A. 6-Phthalimido-1-bromohexane
A solution of phthalic anhydride (32.0 g, 0.21 mol) and 6-aminohexanol
(25.0 g, 0.21 mol) in 180 mL dry toluene was heated to reflux for 3 hrs.
When the intermediate phthalimide was formed, PBr.sub.3 (18.2 mL, 0.58 mol
in 20 mL toluene) was added dropwise and the reaction was stirred at
100.degree. C. for 1.5 hrs. The reaction mixture was filtered thru a
microfiber filter and the filtrate was concentrated in vacuo to give the
crude product which then crystallized from toluene to give
6-phthalimido-1-bromohexane. (86% yield).
B. N-Boc-2-pyrroldinethio-6-hexaphthalimide
To a solution of the Example I Part A tosylate (8 g, 22.53 mmol) in 150 mL
acetone was added KSAc (3.34 g, 29.3 mmol). The reaction was heated to
reflux for 16 hrs. The precipitate was removed by filtration and the
filtrate was concentrated in vacuo. The crude product was isolated by a
silica gel column using a mixture of EtOAc and hexane (2 to 1 ratio) as an
eluting solvent to give N-Boc-2-pyrrolidinethio-acetate (43% yield).
(M+H).sup.+ @ 260.
To a solution of N-Boc-2-pyrrolidinethioacetate (0.9 g, 3.47 mmol) and
6-phthalimido-1-bromohexane (1.08 g, 3.47 mmol) in 10 mL MeOH and 1 mL of
DMF was added NaOMe (0.8 mL, 3.47 mmol) under argon at 0.degree. C. After
the reaction was stirred at 0.degree. C. for 30 minutes, the ice bath was
removed; the reaction was stirred at room temperature for 16 hrs and was
concentrated in vacuo. The crude product was purified on a silica gel
column using EtOAc/Hexane (1 to 4 ratio) to give title compound (35%
yield).
##STR118##
To a solution of N-Boc-2-pyrrolidine thio-6-hexaphthalimide (0.5 g, 11.2
mmol) in 3 mL of CH.sub.2 Cl.sub.2 was added 4N HCl (3.5 mL, 3.5 mmol) at
0.degree. C. The ice bath was removed and the reaction was stirred at room
temperature for 1 hr. The reaction mixture was concentrated in vacuo to
give the title HCl salt (89% yield).
##STR119##
To a solution of 2-naphthalene sulfonyl chloride (3 g, 13.23 mmol) and
serine benzylester (3.07 g, 13.23 mmol) in 120 mL CH.sub.2 Cl.sub.2 was
added Et.sub.3 N (5.53 mL, 13.23 mmol) at 0.degree. C. The reaction was
stirred at room temperature for 1 hr and washed with a sat. solution of
KHSO.sub.4, brine over Na.sub.2 SO.sub.4. The organic layer was
concentrated in vacuo to give title compound (93% yield). (M+H).sup.+ @
386.
D(2). 2-Naphthalenesulfonyl-serine
To a solution of Part D(1) compound (1 g, 2.59 mmol) and Pd/C (10%, 50 mg)
in 25 mL EtOAc was added 2.6 mL of 1N HCl, followed by H.sub.2. The
reaction was stirred at room temperature for 16 hrs and the catalyst was
removed by filtration. The filtrate was concentrated in vacuo to give
2-naphthalene sulfonyl-serine (89% yield). (M+H).sup.+ @ 296.
##STR120##
To a solution of 2-naphthalene sulfonyl-serine (293 mg, 0.99 mmol) and HOBT
(1.47 g, 1.09 mmol) in 10 mL DMF was added WSC (209 mg, 1.09 mmol) and
Part C compound (380 mg, 0.99 mmol), followed by NMM (0.3 mL, 2.18 mmol).
The reaction was stirred at room temperature for 16 hrs. The reaction
mixture was partitioned between EtOAc and a sat. solution of NaHCO.sub.3
and the organic layer was washed with a sat. solution of KHSO.sub.4,
brine, dried over Na.sub.2 SO.sub.4, and EtOAc was concentrated in vacuo
to give title compound (83% yield).
##STR121##
To a solution of Part D compound (510 mg, 0.82 mmol) in 10 mL MeOH was
added hydrazine (0.26 mL, 8.2 mmol) at room temperature. The reaction was
stirred at 40.degree.-50.degree. C. for 4 hrs. The reaction mixture was
concentrated in vacuo and partitioned between EtOAc and 1N NaOH, dried
over Na.sub.2 SO.sub.4. The organic layer concentrated in vacuo to give
title compound (74% yield).
F.
S-(R*,R*)!-N-2-2-6-(Aminoiminomethyl)amino!hexyl!thio!methyl!-1-py
rrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
trifluoroacetate
To a solution of Part E amine (300 mg, 0.61 mmol) and amidinesulfonic acid
(113 mg, 0.91 mmol) in 10 mL EtOH was added Et.sub.3 N (0.26 mL, 1.83
mmol). The reaction stirred at room temperature for 16 hrs. The reaction
mixture was concentrated in vacuo and purified by a preparative HPLC to
give title compound (34% yield).
Elemental Analysis: C.sub.25 H.sub.37 N.sub.5 O.sub.4 S.1.15TFA.0.52
H.sub.2 O Calc.: C, 47.15; H, 5.71; N, 10.44; S, 9.56 Found: C, 47.15; H,
5.42; N, 10.36; S, 9.91.
.alpha.!.sub.D =-91.0 (c=2.00, CH.sub.3 OH).
EXAMPLE XIII
3R-3R*,3(S*,S*)!!-N-2-2-1-(Aminoiminomethyl)-3-piperidinyl!methyl!t
hio!methyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulf
onamide, trifluoroacetate (1:1) salt
A. N-Carbobenzyloxy-3-piperidinemethanol
To a solution of 3(R)-N-carbobenzyloxy-3-carboxypiperidine (2.0 g, 7.6
mmol) in 15 mL THF was added BH.sub.3.Me.sub.2 S dropwise at 0.degree. C.
and the ice bath was removed. The reaction was stirred at room temperature
for 2 hrs. The reaction mixture was quenched with HOAc, MeOH, then 1N 7.6
mL NaOH and was stirred at room temperature for 2 hrs. The reaction
mixture was concentrated in vacuo and extracted with EtOAc. The EtOAc
layer was washed with 1N NaOH and concentrated in vacuo to give
N-carbobenzyloxy-3-piperidinemethanol.
##STR122##
To a solution of Part A compound (2.0 g, 8.03 mmol) and TsCl (3.06 g, 16.06
mmol) in 35 mL CH.sub.2 Cl.sub.2 was added Et.sub.3 N (3.39 mL, 24.09
mmol) at 0.degree. C. The ice bath was removed and the reaction was
stirred at room temperature for 16 hrs. The reaction mixture was washed
with H.sub.2 O and was concentrated in vacuo to give the crude which was
purified by a silica gel column using a mixed solvent (EtOAc:hexane=1:2
ratio) to obtain title compound (70% yield).
##STR123##
To a solution of N-Boc-2-pyrrolidinethioacetate (Example XII Part B) (1.0
g, 3.86 mmol) in 30 mL DMF was added NaOMe (0.88mL, 15.44 mmol) under
argon at 0.degree. C. The reaction was stirred at 0.degree. C. for 5
minutes and Part B compound (1.5 g, 3.86 mmol) was added at 0.degree. C.
The ice bath was removed and stirred at room temperature for 2 hrs. The
reaction mixture was partitioned between EtOAc and H.sub.2 O and the EtOAc
was washed with brine, dried over Na.sub.2 SO.sub.4, concentrated in vacuo
to give title compound (79% yield).
##STR124##
To a solution of Part C compound (1.2 g, 2.46 mmol) in 2 mL CH.sub.2
Cl.sub.2 was added 4N HCl (7.4 mL, 7.4 mmol) at 0.degree. C. The ice bath
was removed and the reaction was stirred at room temperature for 2 hrs.
The reaction was concentrated in vacuo to give title compound.
##STR125##
To a solution of 2-naphthalene sulfonyl-serine (prepared in Example XII
Part D) (0.78 g, 2.67 mmol) and HOBT (0.4 g, 2.93 mmol) in 11 mL DMF was
added WSC (0.56 g, 2.93 mmol), followed by Part D compound (1.03 g, 2.67
mmol). After 5 minutes, NMM (0.82 mL, 5.86 mmol) was added to adjust the
pH to 7.5-8.0 and the reaction stirred at room temperature for 16 hrs. The
reaction mixture was partitioned between EtOAc and a sat. solution of
NaHCO.sub.3 and the EtOAc layer was washed with a sat. solution of
KHSO.sub.4, brine, dried over Na.sub.2 SO.sub.4. The organic layer was
concentrated in vacuo to give title compound (87% yield).
##STR126##
To a solution of Part E compound (1.3 g, 2.08 mmol) in 3 mL CH.sub.2
Cl.sub.2 was added HBr (6.3 mL, 6.3 mmol) at 0.degree. C. The ice bath was
removed and the reaction was stirred at room temperature for 1 hr. The
reaction mixture was concentrated in vacuo to give the title HBr salt (98%
yield).
##STR127##
To a solution of the Part F piperidine HBr salt (1.08 g, 2.04 mmol) and
H-pyrazole1-carboxamidine.HCl (490 mg, 3.34 mmol) in 10 mL DMF was added
diisopropylethylamine (DIEA) (1.06 mL, 6.08 mmol) and the reaction was
stirred at room temperature for 72 hrs. The reaction mixture was purified
by a preparative HPLC to give title compound (27% yield).
H.
3R-3R*,3(S*,S*)!!-N-2-2-1-(Aminoiminomethyl)-3-piperidinyl!methyl!
thio!methyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesul
fonamide, trifluoroacetate (1:1) salt
##STR128##
To a solution of Part G compound (0.31 g, 0.54 mmol) in 3 mL MeOH was added
1N NaOH (1.62 mL, 1.62 mmol) at 0.degree. C. The ice bath was removed and
the reaction was stirred at room temperature for 2 hrs. The reaction
mixture was concentrated in vacuo and purified by preparative HPLC to give
title compound (97% yield).
Elemental Analysis: C.sub.25 H.sub.35 N.sub.5 O.sub.4 S.sub.2.1.4 TFA.1.25
H.sub.2 O Calc.: C, 46.64; H, 5.48; N, 9.78; S, 8.96; F, 11.15 Found: C,
46.64; H, 5.28; N, 9.77; S, 8.77; F, 11.16.
.alpha.!.sub.D =-49.3.degree. (c=4.1, CD.sub.3 OD).
EXAMPLE XIV
3S-3R*,3(R*,R*)!!-N-2-2-1-(Aminoiminomethyl)-3-piperidinyl!methyl!t
hio!methyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulf
onamide, trifluoroacetate (1:1) salt
##STR129##
Following the procedure of Example XIII, except substituting
3(S)-N-carbobenzyloxy-3-carboxypiperidine for the 3(R) isomer used in Part
A of Example XIII, the title compound was obtained.
Elemental Analysis: C.sub.25 H.sub.35 N.sub.5 O.sub.4
S.sub.2.1.15TFA.1.20H.sub.2 O Calc.: C, 47.77; H, 5.66; N, 10.20; S, 9.34;
F, 9.55 Found: C, 47.77; H, 5.37; N, 10.31; S, 8.99; F, 9.57.
.alpha.!.sub.D =-21.0.degree. (c=4.08, DMSO).
Examples of additional compounds which may be prepared following the
procedures set out in Examples I to XIV and in the specification are set
out below.
__________________________________________________________________________
##STR130##
Example
R.sup.6' R R.sup.2
R.sup.1
Xa R.sub.a n
__________________________________________________________________________
XV
##STR131## HOCH.sub.2 H CH.sub.3
S
##STR132## 0
XVI
##STR133##
##STR134## H CH.sub.3
SO
##STR135## 1
XVII
##STR136## H CH.sub.3 S
H SO.sub.2
##STR137## 2
XVIII
H
##STR138## CH.sub.3 O
##STR139##
S
##STR140## 2
XIX
##STR141## NH.sub.2 COCH.sub.2 CH.sub.2
OH
##STR142##
SO
##STR143## 0
XX
##STR144## CH.sub.3 CH(OH)
H
##STR145##
SO.sub.2
##STR146## 1
XXI
##STR147## HO.sub.2 CCH.sub.2 CH.sub.2
H NH.sub.2
S
##STR148## 1
XXII C.sub.4 H.sub.9 CO.sub.2
##STR149##
##STR150##
SO
##STR151##
__________________________________________________________________________
2
__________________________________________________________________________
The following Examples represent preferred embodiments of the third
embodiment of the present invention. unless otherwise indicated, all
temperatures are expressed in degrees Centigrade.
EXAMPLE 1a
S-(R*,R*)!-4-Amino-N-1-3-hydroxy-2-(2-naphthalenylsulfonyl)amino!-1-ox
opropyl!-2-pyrrolidinyl!methyl!butanamide, hydrochloride
A. (S)-2-(Azidomethyl)1-pyrrolidinecarboxylic acid, 1,1-dimethylethyl ester
(S)-2-(4-Methylphenyl)sulfonyl!oxy!methyl!-1-pyrrolidinecarboxylic acid,
1,1-dimethylethyl ester (3.55 g, 10 mmol, see J. Das et al, J. Med Chem,
1992, Vol. 35,2610 for preparation) was dissolved in dimethyl sulfoxide
(DMSO) (40 mL) and treated with sodium azide (980 mg, 15 mmol). The
mixture was heated at 70.degree. C. for 5 hours, then cooled and diluted
with ether. After washing with water (3.times.50 mL), the ether layer was
dried (MgSO.sub.4) and freed of solvent in vacuo to give title compound as
a colorless oil (2.05 g, 91%) which was used without further purification.
B. N-BOC-L-Proline Methylamine
The Part A azide (1.45 g, 6.4 mmol) was dissolved in absolute ethanol (50
mL), and treated with 10% palladium on carbon (290 mg). The flask was
equipped with a hydrogen filled balloon via a three way stopcock. Air
inside the flask was evacuated under reduced pressure and the flask was
then filled with hydrogen from the balloon. This process was repeated
three times. The mixture was stirred overnight in the hydrogen atmosphere.
The catalyst was then removed by filtration through a pad of MgSO.sub.4
and the pad was washed with more ethanol. The solvent was removed in vacuo
to give title compound as a colorless oil (1.19 g, 93%).
C.
(S)-2-1-Oxo-4-(phenylmethoxy)carbonyl!amino!butyl!amino!methyl!-1-pyr
rolidinecarboxylic acid, 1,1-dimethylethyl ester
Part B N-BOC-L-proline methylamine (1.04 g, 5.2 mmol) and
N-CBZ-4-aminobutyric acid (1.24 g, 5.2 mmol) were dissolved in dimethyl
formamide (DMF) (20 mL) at room temperature (RT). HOBT (702 mg, 5.2 mmol),
WSC (995 mg, 5.2 mmol) and NMM (570 .mu.L, 5.2 mmol) were added. The
reaction was stirred for 16 hours (h), partitioned between ethyl
acetate(50 mL) and 10% KHSO.sub.4 (100 mL). The aqueous layer was
extracted with ethyl acetate (2.times.50 mL), and the organic layers were
combined. The combined organic layers were washed with saturated
NaHCO.sub.3 (50 mL) and saturated NaCl (50 mL), dried over magnesium
sulfate and concentrated in vacuo to provide an oil. The crude material
was purified by chromatography on silica gel. Elution with ethyl
acetate:hexanes(2:1) followed by ethyl acetate gave title compound, (1.809
g, 83%) as a viscous oil which was characterized by NMR and used in the
next step.
D.
(S)-4-(Phenylmethoxy)carbonyl!amino!-N-(2-pyrrolidinylmethyl)butanamide,
trifluoroacetate salt
Part C BOC derivative (915 mg, 2.19 mmol) was dissolved in trifluoroacetic
acid (TFA) (15 mL) and stirred at RT 80 min. The TFA was removed by
distillation under reduced pressure and by coevaporation with toluene to
give title amine.
E.
S-(R*,R*)!-2-Oxo-2-2-1-oxo-4-(phenylmethoxy)carbonyl!amino!butyl!a
mino!methyl!-1-pyrrolidinyl!-1-(phenylmethoxy)methyl!ethyl!carbamic acid,
1,1-dimethylethyl ester
The Part D crude TFA salt (2.19 mmol) and BOC-Ser(OBn)-OH (710 mg, 2.41
mmol) were dissolved in DMF (10 mL) at RT. HOBT (326 mg, 2.41 mmol), WSC
(463 mg, 2.41 mmol) and NMM (570 .mu.L) were added. The reaction was
stirred for 18 h, partitioned between ethyl acetate (50 mL) and saturated
KHSO.sub.4 solution (50 mL). The aqueous layer was reextracted with ethyl
acetate (2.times.50 mL) and the organic layers were combined. The combined
organic layers were washed with saturated NaHCO.sub.3 (30 mL) and
saturated NaCl, dried over magnesium sulfate and concentrated in vacuo.
The remaining yellow oil was purified by chromatography on silica gel.
Elution with ethyl acetate gave title compound, (611 mg, 47%) which was
characterized by NMR.
F.
S-(R*,R*)!-N-1-2-(2-Naphthalenylsulfonyl)amino!-1-oxo-3-(phenylmethox
y)propyl!-2-pyrrolidinyl!methyl!-4-(phenylmethoxy)carbonyl!amino!butanami
de
Part E BOC derivative (610 mg, 1.02 mmol) was dissolved in TFA (15 mL) and
stirred at RT 100 min. The TFA was removed by distillation under reduced
pressure and by coevaporation with toluene. The crude TFA salt was
dissolved in dichloromethane (15 mL), cooled in an ice bath, and
2-naphthylsulfonyl chloride (Aldrich, 254 mg, 1.12 mmol) was added,
followed by triethylamine (855 .mu.L, 6.12 mmol). The solution was warmed
to RT and stirred 1.75 h before diluting with dichloromethane (50 mL).
This solution was washed with potassium hydrogen sulfate solution
(2.times.30 mL) and saturated NaHCO.sub.3 (2.times.30 mL), dried over
magnesium sulfate, and evaporated to provide crude title compound. The
crude product was chromatographed on silica gel and eluted with
EtOAc:hexanes (2:1) followed by EtOAc followed by 5%MeOH in CH.sub.2
Cl.sub.2 to provide title sulfonamide as a foam (384 mg, 72% overall in
two steps).
G.
S-(R*,R*)!-4-Amino-N-1-3-hydroxy-2-(2-naphthalenylsulfonyl)amino!-1-o
xopropyl!-2-pyrrolidinyl!methyl!butanamide, hydrochloride
Part F compound (480 mg, 0.71 mmol) was dissolved in ethanol (60 mL) to
which acetyl chloride (1.4 mL) had been added and the mixture was
hydrogenated over 10% Pd--C (150 mg) at 55 psi. After 17 h, additional
catalyst (100 mg) and acetyl chloride (0.5 mL) were added. After an
additional 18 h, the reaction mixture was filtered and the residual
catalyst was washed with EtOH. The filtrate was concentrated in vacuo to
obtain title des-benzyl, des-CBZ product.
EXAMPLE 2a
S-(R*,R*)!-4-(Aminoiminomethyl)amino!-N-1-3-hydroxy-2-(2-naphthalenyl
sulfonyl)amino!-1-oxopropyl!-2-pyrrolidinyl!methyl!butanamide,
trifluoroacetate (1:1) salt
Example 1a compound (0.71 mmol) was dissolved in ethanol (20 mL), to which
amidinesulfonic acid (124 mg, 1.0 mmol) and triethylamine (270 .mu.L, 1.9
mmol) were added. After 2.5 h the mixture was filtered through a pad of
celite, washed with ethanol and the filtrate was concentrated to dryness.
The crude material was purified by preparative HPLC to provide a white
solid (218 mg, 47% overall yield from Example 1, Part F),
Purity.gtoreq.98%.
.alpha.!.sub.D =-37.2.degree. (c=0.9, MeOH)
Analysis calcd for 1.25 TFA+0.60 H.sub.2 O: C, 46.55; H, 5.28; N, 12.77; F,
10.83; S, 4.87. Found: C, 46.54; H, 5.56; N, 12.69; F, 10.80; S, 4.93.
EXAMPLE 3a
R-(R*,S*)!-2-2-(4-Aminobutyl)amino!methyl!-1-pyrrolidinyl!-2-oxo-1-(ph
enylmethyl)ethyl!carbamic acid, phenylmethyl ester
A. (S)-2-(Trifluoroacetyl)amino!methyl!-1-pyrrolidinecarboxylic acid,
1,1-dimethylethyl ester
A solution of Example 1a, Part B N-BOC-L-prolinemethylamine compound (2.64
g, 13.27 mmol) in dichloromethane (50 mL) and pyridine (10 mL) was cooled
to 0.degree. C. and treated dropwise with trifluoroacetic anhydride (3.78
mL, 26.55 mmol). The reaction was allowed to warm slowly to room
temperature and left stirring overnight. The mixture was diluted with
ethyl acetate (150 mL) and washed with satd. copper sulfate solution
(5.times.50 mL) and brine (3.times.30 mL), dried (MgSO.sub.4) and freed of
solvent in vacuo to give title compound as a yellow oil (3.516 g, 84%)
which was used without further purification.
B.
(S)-2-4-(1,3-Dihydro-1,3-dioxo-1H-isoindol-2-yl)butyl!(trifluoroacetyl)
amino!methyl!-1-pyrrolidinecarboxylic acid, 1,1-dimethylethyl ester
The Part A trifluoroacetamide (3.514 g, 11.12 mmol) and
N-(4-bromobutyl)-phthalimide (3.136 g, 11.12 mmol) were dissolved in DMF
(60 mL), and treated with cesium carbonate (7.25 g, 22.24 mmol). The
reaction was heated in an oil bath maintained at 55.degree. C. for 22
hours, cooled to room temperature and partioned between ethyl acetate (150
mL) and water (100 mL). The layers were separated, and the aqueous layer
was reextracted with ethyl acetate (50 mL). The combined organic layers
were dried over magnesium sulfate and concentrated in vacuo. The crude
material was purified by chromatography on silica gel (Merck, 400 mL),
eluting with 20-40% ethyl acetate in hexane to give title compound (2.773
g, 50%) as a viscous foam.
C.
R-(R*,S*)!-2-2-4-(1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)butyl!(trif
luoroacetyl)amino!methyl!-1-pyrrolidinyl!-2-oxo-1-(phenylmethyl)ethyl!carba
mic acid, phenylmethyl ester
Part A BOC derivative (2.27 g, 4.567 mmol) was dissolved in dichloromethane
(10 mL) and TFA (30 mL) and stirred at RT 2.5 hours. The TFA was removed
by distillation under reduced pressure and by coevaporation with toluene
to give the TFA salt of the amine. This and Z-D-phenylalanine (1.50 g,
5.02 mmol) were dissolved in DMF (20 mL) at RT. HOBT (680 mg, 5.02 mmol),
WSC (965 mg, 5.02 mmol) and NMM (1.1 mL, 10 mmol) were added. The reaction
was stirred for 16 h, partitioned between ethyl acetate (70 mL) and 10%
KHSO.sub.4 (70 mL). The aqueous layer was extracted with ethyl acetate
(2.times.70 mL), and the organic layers were combined. The combined
organic layers were washed with saturated NaHCO.sub.3 (50 mL) and
saturated NaCl (50 mL), dried over magnesium sulfate and concentrated in
vacuo to provide a viscous oil. The crude material was purified by
chromatography on silica gel. Elution with ethyl acetate:hexanes(1:1)
followed by ethyl acetate gave title phthalimide, (2.43 g, 78%) as a
viscous oil which was characterized by NMR.
D.
R-(R*,S*)!-2-2-(4-Aminobutyl)amino!methyl!-1-pyrrolidinyl!-2-oxo-1-(p
henylmethyl)-ethyl!carbamic acid, phenylmethyl ester
Part C phthalimide (1.79 g, 2.63 mmol) was dissolved in ethanol (12 mL) and
treated with methyl hydrazine (2 mL). The mixture was heated under reflux
for 2 hours at which time the HPLC indicated that no starting material
remained, but showed a mixture of products. The solvent was removed in
vacuo and by co-evaporation with toluene. The residue was dissolved in
methanol (50 mL) and water (15 mL) and treated with potassium carbonate (2
g). The mixture was left stirring overnight at room temperature to
complete the removal of the trifluoroacetamide group. 1N HCl solution was
added to acidify the mixture and non-basic organics were removed by
extracting with dichloromethane (70 mL). The dichloromethane layer was
extracted with 1N HCl (50 mL). The combined aqueous layers were basified
with NaOH solution and the desired amine was extracted into
dichloromethane (3.times.70 mL), dried over magnesium sulfate and
concentrated in vacuo to provide title compound as an oil (1.065 g, 90%).
EXAMPLE 4a
R-(R*,S*)!-2-2-4-(Aminoiminomethyl)amino!butyl!amino!methyl!-1-pyrro
lidinyl!-2-oxo-1-(phenylmethyl)ethyl!carbamic acid, phenylmethyl ester,
trifluoroacetate (1:2) salt
Example 3a compound (1.069 g, 2.356 mmol) was dissolved in ethanol (60 mL),
to which amidinesulfonic acid (352 mg, 2.84 mmol) and triethylamine (550
.mu.L, 3.94 mmol) were added. After 3.5 h the mixture was filtered through
a pad of celite, washed with ethanol and the filtrate was concentrated to
dryness. The crude material was purified by preparative HPLC to provide a
white solid (1.225 g, 69%), Purity.gtoreq.98%.
.alpha.!.sub.D =-2.9.degree. (c=0.5 MeOH)
Analysis calcd for 2.20TFA+0.70 H.sub.2 O: C, 49.75; H, 5.53; N, 11.09; F,
16.54. Found: C, 49.71; H, 5.48; N, 11.09; F, 16.57.
EXAMPLE 5a
(2S)-N-4-1-(D-Phenylalanyl)-2-pyrrolidinyl!methyl!amino!butyl!guanidine
, trifluoroacetate (1:2) salt
Example 4a compound (540 mg, 0.712 mmol) was dissolved in methanol (30
mL)and treated with Periman's catalyst (Pd(OH).sub.2) (125 mg). The flask
was equipped with a hydrogen filled balloon via a three-way stopcock. Air
inside the flask was removed under reduced pressure and replaced with
hydrogen from the balloon. This process was repeated three times. The
mixture was stirred under the hydrogen atmosphere for 2.5 hours. The
catalyst was removed by filtration and the filter pad was washed with
additional methanol. The filtrate was freed of solvent in vacuo to give
title compound which was dissolved in water (30 mL), passed through a
millipore membrane and lyophilized to provide title compound as a white
solid (429 mg, 93%), Purity.gtoreq.98%.
Analysis calcd for 2.50 TFA+0.20 H.sub.2 O: C, 44.41; H, 5.72; N, 12.95; F,
21.95. Found: C, 44.49; H, 5.70; N, 12.86; F, 21.91.
EXAMPLE 6a
R-(R*,S*)!-2-2-(4-Aminobutyl)sulfonyl!amino!methyl!-1-pyrrolidinyl!-2
-oxo-1-(phenylmethyl)ethyl!carbamic acid, 1,1-dimethylethyl ester
A. 4-Bromobutylsulfonylchloride
A stirred suspension of anhydrous sodium bromide (1.13 g, 11 mmol) and
1,4-butane sulfone (1.36 g, 10 mmol) in DMF (10 mL) was heated in a bath
maintained at 80.degree. C. for 2 hours. The clear solution was cooled to
room temperature and a white solid precipitated. The mixture was diluted
with ethyl acetate and filtered. The white solid collected was washed five
times with ether and dried in vacuo to give the sodium salt of
4-bromobutyl sulfonic acid (2.18 g). Solid phosphorous pentachloride (3.12
g, 15 mmol) was added to the sodium salt in several portions with mixing.
After addition, the solid mixture was triturated with a spatula. An
exothermic reaction ensued, the mixture became colored and became fluid.
After stirring at room temperature for 30 min., the mixture was diluted
with dichloromethane and treated slowly with ice water. The layers were
separated and the dichloromethane solution was dried (MgSO.sub.4),
filtered and the solvent was removed in vacuo. The crude colorless oil was
purified on silica gel (Merck) eluting with 50% ethyl acetate in hexane to
give title compound as a colorless oil (1.69 g, 72%).
B.
R-(R*,S*)!-2-2-(Aminomethyl)-1-pyrrolidinyl!-2-oxo-1-(phenylmethyl)ethy
l!carbamic acid, 1.1-dimethylethyl ester
(1)
R-(R*,S*)!-2-2-(Azidomethyl)-1-pyrrolidinyl!-2-oxo-1-(phenylmethyl)ethy
l!carbamic acid, 1,1-dimethylethyl ester
Example 1a, Part A azide (4.52 g, 20 mmol) was dissolved in dichloromethane
(5 mL) and trifluoroacetic acid (10 mL) and stirred at RT 2 hours. The TFA
was removed by distillation under reduced pressure and by coevaporation
with toluene to give the amine as a TFA salt. This and
t-BOC-D-phenylalanine (5.3 g, 20 mmol) were dissolved in DMF (30 mL). HOBT
(2.7 g, 20 mmol), WSC (4 g, 20 mmol) and NMM (8 mL) were added. The
reaction was stirred for 24 h, diluted with ethyl acetate(75 mL) and
washed with 10% KHSO.sub.4 (2.times.), saturated sodium bicarbonate
solution (2.times.) and 10% lithium chloride solution (2.times.). The
ethyl acetate layer was dried over magnesium sulfate and concentrated in
vacuo to provide title compound (7.4 g, 99%) as an oil which was used
without further purification.
(2)
R-(R*,S*)!-2-2-(Aminomethyl)-1-pyrrolidinyl!-2-oxo-1-(phenylmethyl)ethy
l!carbamic acid, 1,1-dimethylethyl ester
The Part (1) azide (2.25 g, 6.0 mmol) was dissolved in absolute ethanol (40
mL), and treated with 10% palladium on carbon (200 mg). The flask was
equipped with a hydrogen filled balloon via a three way stopcock. Air
inside the flask was evacuated under reduced pressure and the flask was
then filled with hydrogen from the balloon. This process was repeated
three times. The mixture was stirred overnight in the hydrogen atmosphere.
The catalyst was then removed by filtration through a pad of MgSO.sub.4
and the pad was washed with more ethanol. The solvent was removed in vacuo
to give title amine as a white foam (2.15 g, Quant).
C.
R-(R*,S*)!-2-2-(4-Bromobutyl)sulfonyl!amino!methyl!-1-pyrrolidinyl!-
2-oxo-1-(phenylmethyl)ethyl!carbamic acid, 1,1-dimethylethyl ester
A solution of Part A 4-bromobutyl sulfonyl chloride (942 mg, 4 mmol) in
dichloromethane (3 mL) was added dropwise to a cooled (-10.degree. C.)
stirred solution of Part B amine (868 mg, 2.5 mmol) and triethyl amine
(1.2 mL, 8 mmol) in dichloromethane (10 mL). After a few minutes, a
precipitate formed. The suspension was stirred at -10.degree. C. for 2
hours, diluted with dichloromethane (25 mL) and washed with 1N HCl
solution (2.times.15 mL). The dichloromethane solution was dried
(MgSO.sub.4), filtered and the solvent was removed in vacuo. The crude oil
was purified on silica gel (Merck) eluting with 30%, 50%, and 70% ethyl
acetate in hexane to give title compound as a white foam (910 mg, 67%).
D.
R-(R*,S*)!-2-2-(4-Azidobutyl)sulfonyl!amino!methyl!-1-pyrrolidinyl!-
2-oxo-1-(phenylmethyl)ethyl!carbamic acid, 1,1-dimethylethyl ester
A solution of Part Chromosulfonamide (800 mg, 1.46 mmol) in DMSO (5 mL) was
treated with sodium azide (143 mg, 2.2 mmol) and the mixture was heated in
an oil bath maintained at 60.degree. C. for 3.5 hours. After cooling, the
mixture was partitioned between ether (50 mL) and water (25 mL). The ether
layer was washed with water (2.times.10 mL), dried (MgSO.sub.4), filtered
and the solvent was removed in vacuo. The crude oil was purified on silica
gel (Merck) eluting with 50% ethyl acetate in hexane to give title
compound as an oil (733 mg, 98%).
E.
R-(R*,S*)!-2-2-(4-Aminobutyl)sulfonyl!amino!methyl!-1-pyrrolidinyl!-
2-oxo-1-(phenylmethyl)ethyl!carbamic acid, 1,1-dimethylethyl ester
Part D compound (730 mg, 1.437 mmol) was dissolved in ethanol (20 mL)and
treated with 10% palladium on carbon (180 mg). The flask was equipped with
a hydrogen filled balloon via a three-way stopcock. Air inside the flask
was removed under reduced pressure and replaced with hydrogen from the
balloon. This process was repeated three times. The mixture was stirred
under the hydrogen atmosphere for 18 hours. The catalyst was removed by
filtration and the filter pad was washed with additional ethanol. The
filtrate was taken to dryness in vacuo leaving title compound as a
colorless oil (758 mg, Quant.).
EXAMPLE 7a
R-(R*,S*)!-4-(Aminoiminomethyl)amino!-N-1-(2-amino-1-oxo-3-phenylpropyl
)-2-pyrrolidinyl!methyl!butanesulfonamide, trifluoroacetate (1:2) salt
Example 6a amine (1.437 mmol) was dissolved in ethanol (30 mL), to which
amidinesulfonic acid (250 mg, 2.0 mmol) and triethyl amine (280 .mu.L, 2.0
mmol) were added. After 3 h, the mixture was filtered through a pad of
celite, washed with ethanol and the filtrate was concentrated to dryness.
The residue was treated with TFA (5 mL) for 3 hours and then freed of
excess TFA by coevaporation with toluene. The crude material was purified
by preparative HPLC to provide a white solid (523 mg, 55% from Example 6,
Part D compound), Purity.gtoreq.98%.
.alpha.!.sub.D =-63.3.degree. (c=0.6, MeOH).
Analysis calcd for 2.2 TFA+0.2 H.sub.2 O: C, 41.39; H, 5.14; N, 12.38; F,
18.47; S, 4.72. Found: C, 41.44; H, 5.16; N, 11.89; F, 18.41; S, 4.56.
EXAMPLE 8a
R-(R*,S*)!-2-2-(3-Aminopropyl)sulfonyl!amino!methyl!-1-pyrrolidinyl!-
2-oxo-1-(phenylmethyl)ethyl!carbamic acid, 1,1-dimethylethyl ester
A. 3-Bromopropyl sulfonyl chloride
A stirred suspension of anhydrous sodium bromide (1.60 g, 15.5 mmol) and
1,3-propane sulfone (1.83 g, 15 mmol) in DMF (15 mL) was heated in a bath
maintained at 80.degree. C. for 2 hours, during this time material
precipated. The mixture was cooled to room temperature, dilited with ethyl
acetate and filtered. The white solid collected was washed five times with
ether and dried in vacuo to give the sodium salt of 3-bromopropyl sulfonic
acid (3.11 g). Solid phosphorous pentachloride (4.68 g, 22.5 mmol) was
added to the sodium salt, in several portions, with mixing. After
addition, the solid mixture was triturated with a spatula. An exothermic
reaction ensued, the mixture became colored and became fluid. After
stirring at room temperature for 30 min., the mixture was diluted with
dichloromethane and treated slowly with ice water. The layers were
separated and the dichloromethane solution was dried (MgSO.sub.4),
filtered and the solvent was removed in vacuo. The crude colorless oil was
purified on silica gel (Merck) eluting with 30% ethyl acetate in hexane to
give title compound as a colorless oil (2.544 g, 76%).
B.
R-(R*,S*)!-2-2-(3-Bromopropyl)sulfonyl!amino!methyl!-1-pyrrolidinyl!
-2-oxo-1-(phenylmethyl)ethyl!carbamic acid, 1.1-dimethylethyl ester
A solution of Part A 3-bromopropyl sulfonyl chloride (1.06 g, 4.8 mmol) in
dichloromethane (5 mL) was added dropwise to a cooled (-10.degree. C.)
stirred solution of Example 6a, Part B amine (1.05 g, 3.02 mmol) and
triethyl amine (1.44 mL, 9.6 mmol) in dichloromethane (15 mL), After a few
minutes, a precipitate formed. The suspension was stirred at -10.degree.
C. for 2.5 hours, diluted with dichloromethane (35 mL) and washed with 1N
HCl solution (2.times.15 mL). The dichloromethane solution was dried
(MgSO.sub.4), filtered and the solvent was removed in vacuo. The crude oil
was purified on silica gel (Merck, 150 mL) eluting with 50% ethyl acetate
in hexane to give title compound as a white foam (1.160 g, 72%).
C.
R-(R*,S*)!-2-2-(3-Azidopropyl)sulfonyl!amino!methyl!-1-pyrrolidinyl!
-2-oxo-1-(phenylmethyl)ethyl!carbamic acid, 1,1-dimethylethyl ester
A solution of Part B bromo sulfonamide (1.10 g, 2.06 mmol) in DMSO (8 mL)
was treated with sodium azide (202 mg, 3.1 mmol) and the mixture was
heated in an oil bath maintained at 60.degree. C. for 3.75 hours. After
cooling, the mixture was partitioned between ether (60 mL) and water (30
mL). The ether layer was washed with water (2.times.15 mL), dried
(MgSO.sub.4), filtered and the solvent was removed in vacuo. The crude oil
was purified on silica gel (Merck, 150 mL) eluting with 50% ethyl acetate
in hexane to give title compound as a white foam (872 mg, 86%).
D.
R-(R*,S*)!-2-2-(3-Aminopropyl)sulfonyl!amino!methyl!-1-pyrrolidinyl!
-2-oxo-1-(phenylmethyl)ethyl!carbamic acid, 1,1-dimethylethyl ester
Part C compound (870 mg, 1.76 mmol) was dissolved in ethanol (25 mL)and
treated with 10% palladium on carbon (200 mg). The flask was equipped with
a hydrogen filled balloon via a three-way stopcock. Air inside the flask
was removed under reduced pressure and replaced with hydrogen from the
balloon. This process was repeated three times. The mixture was stirred
under the hydrogen atmosphere for 18 hours. The catalyst was removed by
filtration and the filter pad was washed with additional ethanol. The
filtrate was taken to dryness in vacuo leaving title compound as a
colorless oil.
EXAMPLE 9a
R-(R*,S*)!-2-2-3-(Aminoiminomethyl)amino!propyl!sulfonyl!amino!meth
yl!-1-pyrrolidinyl!-2-oxo-1-(phenylmethyl)ethyl!carbamic acid,
1,1-dimethylethyl ester, trifluoroacetate (1:1) salt
Example 8a amine (1.76 mmol) was dissolved in ethanol (35 mL) and
amidinesulfonic acid (306 mg, 2.46 mmol) and triethylamine (345 .mu.L, 2.5
mmol) were added. After 3 hours, the mixture was filtered through a pad of
celite, washed with ethanol and the filtrate was concentrated to dryness.
The crude material was purified by preparative HPLC to provide a white
solid (820 mg, 71% from Example 8a, Part C compound), Purity.gtoreq.98%.
.alpha.!.sub.D =-38.4.degree. (c=0.9, MeOH).
Analysis calcd for 1.25 TFA+0.2 H.sub.2 O: C, 46.63; H, 6.09; N, 12.80; F,
10.85; S, 4.88. Found: C, 46.87; H, 6.12; N, 12.60; F, 10.83; S, 4.92.
EXAMPLE 10a
R-(R,,S*)!-3-(Aminoiminomethyl)amino!-N-1-(2-amino-1-oxo-3-phenylpropyl
!-2-pyrrolidinyl!methyl!propanesulfonamide, trifluoroacetate (1:2) salt
Example 9a compound (500 mg, 0,76 mmol) was treated with TFA (5 mL) for 1.5
hours and then freed of excess TFA in vacuo. The residue was dissolved in
water (20 mL) and passed through a Millipore filtration membrane. The
aqueous solution was frozen and lyophillized to give a dense solid. This
was again dissolved in water (20 mL) and relyophillized to give title
compound as a fluffy white solid (479 mg, 93%), Purity.gtoreq.98%.
.alpha.!.sub.D =-60.2.degree. (c=0.6, MeOH).
Analysis calcd for 2.2 TFA+0.4 H.sub.2 O: C, 40.24; H, 4.98; N, 12.57; F,
18.75; S, 4.80. Found: C, 40.21; H, 5.10; N, 12.35; F, 18.94; S, 4.81.
EXAMPLE 11a
R-(R*,S*)!-2-Amino-1-2-4-(aminomethyl)phenyl!amino!methyl!-1-pyrrolidi
nyl!-3-phenyl-1-propanone, trifluoroacetate (1:3) salt
A. (4-Aminophenyl)methyl!carbamic acid, phenylmethyl ester
To a solution of 4-aminobenzenemethanamine (1.5 g, 12.3 mmol) in 20 mL
CH.sub.2 Cl.sub.2 was added benzylchloroformate (1.57 mL, 11 mmol),
followed by triethylamine (Et.sub.3 N) (1.5 mL, 11 mmol) at room
temperature. White percipitates was observed immediately. The reaction was
stirred at room temperature for 10 minutes and removed Et.sub.3 N salt by
filtration. The reaction mixture was concentrated in vacuo and partitioned
between EtOAc and H.sub.2 O. The EtOAc layer was washed with a sat.
solution of KHSO.sub.4, a sat. solution of NaHCO.sub.3, concentrated in
vacuo and subjected to a silica-gel column using CHCl.sub.3 /CH.sub.3
OH=25/1 ratio as an eluting solvent to give title compound (43% yield).
B.
(S)-4-1-(1,1-Dimethylethoxy)carbonyl!-2-pyrrolidinyl!methyl!amino!ph
enyl!methyl!carbamic acid, phenylmethyl ester
To a solution of Example 16a, Part A aldehyde (0.36 g, 1.79 mmol) and Part
A compound (0.46 g, 1.79 mmol) in 3 mL CH.sub.2 Cl.sub.2 CH.sub.2 Cl.sub.2
was slowly added acetic acid (AcOH) (140 .mu.l, 2.33 mmol) at 0.degree.
C., followed by NaB(AcO).sub.3 H. The reaction stirred at room temperature
for 15 minutes and partitioned between EtOAc/hexane (1:1 ratio) and
H.sub.2 O. The organic layer was washed with H.sub.2 O, brine, dried over
Na.sub.2 SO.sub.4 and concentrated in vacuo to give title compound (98%
yield).
C. (S)-4-(2-Pyrrolidinylmethyl)amino!phenyl!methyl!carbamic acid,
phenylmethyl ester, trifluoroacetate (1:1) salt
To a solution of Part B compound (0.8 g, 1.82 mmol) in 5 mL CH.sub.2
Cl.sub.2 was added trifluoroacetic acid (TFA) (5.5 mL, 5.46 mmol) at
0.degree. C. The ice bath was removed and stirred at room temperature for
1 hr. The reaction mixture was concentrated in vacuo and treated with a
sat. HCl/Et.sub.2 O to give title compound (quant. yield).
D.
R-(R*,S*)!-2-(1,1-Dimethylethoxy)carbonyl!amino!-3-phenyl1-2-4-(
phenylmethoxy)amino!methyl!phenyl!amino!methyl!-1-pyrrolidinyl!-1-propanone
To a solution of Boc-D-Phenylalanine (0.55 g, 2.05 mmol) and HOBT (0.28 g,
2.05 mmol) in 4 mL DMF was added WSC (0.39 g, 2.05 mmol), followed by Part
C compound (0.7 g, 1.87 mmol). NMM (226 ml, 2.05 mmol) was used to adjust
the pH to 7.5-8.0. The reaction stirred at room temperature for 16 hrs and
partitioned between EtOAc and H.sub.2 O. The EtOAc layer was further
washed with a sat. solution of NaHCO.sub.3, a sat. solution of KHSO.sub.4,
brine, dried over Na.sub.2 SO.sub.4 and concentrated in vacuo to give
title compound (41% yield).
E.
R-(R*,S*)!-2-Amino-1-2-4-(aminomethyl)phenyl!amino!methyl!-1-pyrrolid
inyl!-3-phenyl1-propanone, trifluoroacetate (1:3) salt
To a solution of Part D compound (0.2 g, 0.34 mmol) in 1 mL HOAc was added
slowly 30% HBr in HOAc (1 mL, 1.12 mmol) at 0.degree. C. The ice bath was
removed and the reaction was stirred at room temperature for 1 hr. The
reaction mixture was concentrated in vacuo and added Et.sub.2 O with
stirred to formed white percipitates which was filtered to give title
compound (59% yield).
Elemental Analysis: C.sub.21 H.sub.28 N.sub.4 O.3.10 TFA.0.60 H.sub.2 O
Calc.: C, 45.58; H, 4.54; N, 7.82; F, 24.65 Found: C, 45.59; H, 4.29; N,
8.09; F, 24.80.
EXAMPLE 12A
R-(R*,S*)!-N-2-2-4-(Aminomethyl)phenyl!amino!methyl!-1-pyrrolidinyl!-
2-oxo-1-(phenylmethyl)ethyl!-2-naphthalenesulfonamide, trifluoroacetate
(2:3) salt
A.
S-(R*,S*)!-2-Amino-3-phenyl1-2-4-(phenylmethoxy)amino!methyl!pheny
l!amino!methyl!-1-pyrrolidinyl!-1-propanone
To a solution of Example 1a, Part D compound (0.2 g, 0.34 mmol) in 2 mL
CH.sub.2 Cl.sub.2 was added TFA (1 mL, 1.12 mmol) at 0.degree. C. The ice
bath was removed and the reaction was stirred at room temperature for 1.5
hrs. The reaction mixture was concentrated in vacuo to give title compound
(97% yield).
B.
R-(R*,S*)!-N-2-2-4-(Phenylmethyl)carbonyl!amino!methyl!phenyl!ami
no!methyl!-1-pyrrolidinyl!-2-oxo-1-(phenylmethyl)ethyl!-2-naphthalenesulfon
amide
To a solution of Part C compound (0.17 g, 0.28 mmol) in 3 mL CH.sub.2
Cl.sub.2 was added 2-naphthalene sulfonyl chloride (0.07 g, 0.31 mmol),
followed by Et.sub.3 N (86 ml, 0.62 mmol). Additional Et.sub.3 N (20 ml,
0.14 mmol) was added after 1 hr. The reaction was stirred at room
temperature for 1 more hr and partitioned between CH.sub.2 Cl.sub.2 and
H.sub.2 O. The CH.sub.2 Cl.sub.2 layer was washed with a sat. solution of
KHSO.sub.4, a sat. solution of NaHCO.sub.3, brine, dried over Na.sub.2
SO.sub.4 and concentrated in vacuo to give title compound (83% yield).
C.
R-(R*,S*)!-N-2-2-4-(Aminomethyl)phenyl!amino!methyl!-1-pyrrolidinyl!
-2-oxo-1-(phenylmethyl)ethyl!-2-naphthalenesulfonamide, trifluoroacetate
(2:3)
To a solution of Part B compound (0.16 g, 0.24 mmol) in 1 mL HOAc was added
30% HBr in HOAc (0.8 mL, 0.71 mmol) at 0.degree. C. The ice bath was
removed and the reaction was stirred at room temperature for 4 hrs. The
reaction mixture was concentrated in vacuo and subjected to a prep. HPLC
using 35 to 90% B over 40 minutes gradient to give title compound (49%
yield).
Elemental Analysis Calcd for C.sub.31 H.sub.34 N.sub.4 O.sub.3 S.1.60 TFA:
C, 56.65; H, 4.95; N, 7.73; F, 12.58; S, 4.42 Found: C, 56.66; H, 4.93; N,
7.79; F, 12.56; S, 4.20.
.alpha.!.sub.D =-35.1 (c=0.51, MeOH).
EXAMPLE 13A
R,(R*,S*)!-2-Amino-1-2-3-(aminoiminomethyl)amino!propyl!-1-pyrrolidinyl
!-2-phenyl-1-propanone, trifluoroacetate (1:3) salt
A.
(S)-2-3-(1,3-Dihydro-1,3-dioxo-1H-isoindol-2-yl)propyl!(trifluoroacetyl
)amino!methyl!-1-pyrrolidinecarboxylic acid, 1,1-dimethylethyl ester
A solution of Example 3a, Part A trifluoroacetamide (1.26 g, 4 mmol) and
N-(3-bromopropyl)phthalimide (1.7 g, 6 mmol) in DMF (40 mL) was treated
with cesium carbonate (2.61 g, 8 mmol). The reaction was heated in an oil
bath maintained at 55.degree. C. for 24 hours, cooled to room temperature
and partitioned between ethyl acetate (75 mL) and water. The layers were
separated. The organic layer was dried over magnesium sulfate and
concentrated in vacuo. The crude material was purified by chromatography
on silica gel (Merck, 400 mL), eluting with 10-30% ethyl acetate in hexane
to give title compound (1.07 g, 53%) as a colorless oil.
B.
R-(R*,S*)!-2-2-3-(1,3-Dihydro-1,3-dioxo-2H-isoindol-2-yl)propyl!(tri
fluoroacetyl)amino!methyl!-1-pyrrolidinyl!-2-oxo-1-(phenylmethyl)ethyl!carb
amic acid, phenylmethyl ester
Part A BOC derivative (974 mg, 2.0 mmol) was dissolved in dichloromethane
(2 mL) and TFA (4 mL) and stirred at RT 2 hours. The TFA was removed by
distillation under reduced pressure and by coevaporation with toluene to
give the TFA salt of the amine as an oil. This and Z-D-phenylalanine (600
mg, 2 mmol) were dissolved in DMF (15 mL) at RT. HOBT (270 mg, 2 mmol),
WSC (382 mg, 2 mmol) and NMM (0.6 mL, 6 mmol) were added. The reaction was
stirred for 16 h, diluted with ethyl acetate (75 mL) and washed with 10%
KHSO.sub.4 (20 mL, 2.times.) and saturated NaHCO.sub.3 (20 mL, 2.times.).
The ethyl acetate extract was dried over magnesium sulfate and
concentrated in vacuo to provide an oil. The crude material was purified
by chromatography on silica gel. Elution with 20, 30, and 50% ethyl
acetate in hexanes gave title compound, (1.06 g, 80%) as a white foam.
C.
R-(R*,S*)!-2-2-(3-Aminopropyl)amino!methyl!-1-pyrrolidinyl!-2-oxo-1-(
phenylmethyl)ethyl!carbamic acid, phenylmethyl ester
Part B phthalimide (1 g, 1.51 mmol) was dissolved in absolute ethanol (15
mL) and treated with methyl hydrazine (1.2 mL). The mixture was heated
under reflux for 2 hours. The solvent was removed in vacuo and by
co-evaporation with toluene (10 mL) and dichloromethane (10 mL.times.2).
The residue was dissolved in methanol (30 mL) and water (10 mL) and
treated with potassium carbonate (1.5 g), The mixture was left stirring
overnight at room temperature to complete the removal of the
trifluoroacetamide group. 1N HCl solution (25 mL) was added to acidify the
mixture and non-basic organics were removed by extracting with
dichloromethane (25 mL, .times.3). The dichloromethane layers were
extracted with 1NHCl (50 mL). The combined aqueous layers were basified
with NaOH solution and the desired amine was extracted into
dichloromethane (30 mL, .times.3), dried over magnesium sulfate and
concentrated in vacuo to provide title compound as an oil (615 mg, 93%).
D.
R-(R*,S*)!-2-2-3-(Aminoiminomethyl)amino!propyl!amino!methyl!-1-pyrr
olidinyl!-2-oxo-1-(phenylmethyl)ethyl!acetate (1:3) salt
Part C compound (615 mg, 1.40 mmol) was dissolved in absolute ethanol (25
mL), to which amidinesulfonic acid (445 mg, 3.5 mmol) and triethylamine
(560 .mu.L, .about.4 mmol) were added and the mixture left stirring 16
hours at room temperature. The mixture was filtered through a pad of
celite. The pad was washed with ethanol and the filtrate was concentrated
to dryness. The crude material was purified by preparative HPLC (YMC S-10
ODS 50.times.500 mm column, eluting with 49-58% methanol in water,
containing 0.1% TFA). Fractions containing title compound were combined
and lyophilized to provide a white solid (620 mg).
E.
R-(R*,S*)!-2-Amino-1-2-3-(aminoiminomethyl)amino!propyl!-1-pyrrolidiny
l!-3-phenyl1-propanone, trifluoroacetate (1.;3) salt
The Part D TFA salt (.about.620 mg, ) was dissolved in methanol (30 mL) and
treated with Pearlman's catalyst (150 mg). The flask was equipped with a
hydrogen filled balloon via a three-way stopcock. Air inside the flask was
removed under reduced pressure and replaced with hydrogen from the
balloon. This process was repeated three times. The mixture was stirred
under the hydrogen atmosphere for 3 hours. The catalyst was removed by
filtration through Celite and the filter pad was washed with additional
methanol. The filtrate was freed of solvent in vacuo to give title
compound which was dissolved in water (20 mL), passed through a millipore
membrane and lyophilized to provide title compound as a white solid (507
mg, 53% from Part C compound), Purity.gtoreq.98%.
Analysis calcd for 2.90 TFA+0.10 H.sub.2 O: C, 42.10; H, 4.91; N, 12.38; F,
24.34. Found: C, 42.10 H, 4.97;, N, 12.28; F, 24.41.
EXAMPLE 14A
S-(R*,S*)!-4-(Aminoiminomethyl)amino!-N-1-(2-amino-1-oxo-3-phenylpropyl
)-2-pyrrolidinyl!methyl!butanamide, trifluoroacetate (1:1) salt
A.
R-(R*,S*)!-N-1-2-(1,1-Dimethylethoxy)carbonyl!amino!-1-oxo-3-phenylp
ropyl!-2-pyrrolidinyl!methyl!-4-(phenylmethoxy)carbonyl!amino!butanamide
Example 6a, Part B compound (694 mg, 2.0 mmol) and Z-4-aminobutyric acid
(478 mg, 2.0 mmol) were dissolved in DMF (10 mL) at RT. HOBT (270 mg, 2.0
mmol), WSC (382 mg, 2.0 mmol) and NMM (500 .mu.L) were added. The reaction
was stirred for 16 h, diluted with ethyl acetate (50 mL) and washed with
10% KHSO.sub.4 (25 mL), saturated NaHCO.sub.3 (25 mL) and saturated NaCl
(30 mL), dried over magnesium sulfate and concentrated in vacuo to provide
title compound (1.04 g, 92%) as a viscous oil which was used without
further purification.
B.
R-(R*,S*)!-4-Amino-N-1-2-(1,1-dimethylethoxy)carbonyl!amino!-1-oxo-3
-phenylpropyl!-2-pyrrolidinyl!methyl!butanamide
Part A compound (1.04 g, 1.83 mmol) was dissolved in ethanol (100 mL) to
which acetyl chloride (150 .mu.L, 2 mmol) had been added and the mixture
was treated with 10% palladium on carbon (400 mg). The flask was equipped
with a hydrogen filled balloon via a three way stopcock. Air inside the
flask was evacuated under reduced pressure and the flask was then filled
with hydrogen from the balloon. This process was repeated three times. The
mixture was stirred in the hydrogen atmosphere for 20 hours. The catalyst
was then removed by filtration and the pad was washed with more ethanol.
The solvent was removed in vacuo to give title compound as a viscous oil
(932 mg, >100%). The material was characterized by NMR and MS. The NMR
indicated an appreciable amount of ethanol was trapped in the product.
C.
R-(R*,S*)!-4-(Aminoiminomethyl)amino!-N-1-2-(1,1-dimethylethoxy)car
bonyl!amino!-1-oxo-3-phenyl-propyl!-2-pyrrolidinyl!methyl!butanamide
Part B BOC derivative (1.83 mmol) was dissolved in ethanol (50 mL), to
which amidinesulfonic acid (318 mg, 2.56 mmol) and triethylamine (690
.mu.L, 4.94 mmol) were added. After 5 h the mixture was filtered through a
pad of celite. The pad was washed with ethanol and the filtrate was
concentrated to dryness to give the title BOC compound which was carried
on without purification.
D.
S-(R*,S*)!-4-(Aminoiminomethyl)amino!-N-1-(2-amino-1-oxo-3-phenylpropy
l)-2-pyrrolidinyl!methyl!butanamide, trifluoroacetate (1:1) salt
Part C BOC derivative (1.83 mmol) was dissolved in TFA (10 mL) and stirred
at RT 90 min. The TFA was removed by distillation under reduced pressure
and by coevaporation with toluene. The crude material was purified by
preparative HPLC (YMC S-10 ODS 30.times.500 mm column, eluting with 38%
methanol in water, containing 0.1% TFA). Fractions containing title
compound were combined and lyophilized to provide a white solid (712 mg,
61% from Part A compound), Purity.gtoreq.98%.
.alpha.!.sub.D =-55.7.degree. (c=0.7, MeOH)
Analysis calcd for 2.25 TFA+0.10 H.sub.2 O: C, 44.60; H, 5.17; N, 13.28; F,
20.26. Found: C, 44.59; H, 5.43; N, 13.17; F, 20.40.
EXAMPLE 15A
S-(R*,S*)!-4-(Aminoiminomethyl)amino!-N-1-(2-(methylsulfonyl)amino!-1-
oxo-3-phenylpropyl!-2-pyrrolidinyl!methyl!butanamide, trifluoroacetate
(1:1) salt
Example 14a compound (158 mg, 0.25 mmol) was dissolved in a mixture of
dichloromethane (2 mL) and dist. THF (4 mL). The solution was treated with
triethyl amine (140 .mu.L) followed by dropwise addition of
methanesulfonyl chloride (24 .mu.L, 0.3 mmol). The mixture was stirred at
room temperature for 4 hours and then treated with water (.about.0.35 mL).
The mixture was taken to dryness in vacuo. The crude material was purified
by preparative HPLC to provide a white solid which was used to take NMR
spectra. The material was recovered, dissolved in water (15 mL), passed
through a millipore membrane and lyophilized to provide title compound as
a white solid (126 mg, 85%).
Analysis calcd for 1.15 TFA+0.60 H.sub.2 O: C, 45.05; H, 5.82; N, 14.14; F,
11.02; S, 5.39. Found: C, 45.34 H, 5.71;, N, 13.75; F, 11.03; S, 5.50.
EXAMPLE 16A
S-(R*,S*)!-4-1-(2-Amino-1-oxo-3-phenylpropyl)-2-pyrrolidinyl!methyl!ami
no!benzenecarboximidamide, trifluoroacetate (1:2) salt
A. (S)-2-Formyl-1-pyrrolidinecarboxylic acid, 1,1-dimethylethyl ester
To a solution of oxalyl chloride (28 mL, 56.2 mmol) in 14 mL dry CH.sub.2
Cl.sub.2 cooled at -78.degree. C. (a dry ice acetone bath) was added 12 mL
dry DMSO over 40 minutes. The reaction was stirred at -74.degree. C. for
additional 40 minutes and (S)-2-(hydroxymethyl)-1-pyrrolidinecarboxylic
acid, 1,1-dimethylethyl ester, (5.40 g, 26.86 mmol) in 14 mL CH.sub.2
Cl.sub.2 was added dropwise at -65.degree. C. over 30 minutes. The
reaction stirred additional 20 minutes at room temperature and partitioned
between Et.sub.2 O and H.sub.2 O. The Et.sub.2 O layer was washed with
more H.sub.2 O (50 mL.times.4) and the H.sub.2 O layers was extracted with
Et.sub.2 O (40 mL.times.3). All the Et.sub.2 O layers was combined, washed
with H.sub.2 O, brine, dried over Na.sub.2 SO.sub.4 and concentrated in
vacuo. The crude product was isolated by a silica-gel column using EtOAc
and hexane (1:5 ratio) as an eluting solvent to give title compound (84%
yield).
B. (S)-2-(4-Cyanophenyl)amino!methyl!-1-pyrrolidinecarboxylic acid,
1,1-dimethylethyl ester
To a solution of Part A aldehyde, (2.0 g, 10.05 mmol) and
4-aminobenzonitrile, (1.3 g, 11.05 mmol) in dichloroethane (10 mL) was
added acetic acid (844 .mu.l, 14.07 mmol) dropwise at 0.degree. C.,
followed by NaB(AcO).sub.3 H (2.34 g, 11.05 mmol). The ice bath was
removed and the reaction stirred at room temperature for 1 hr. The
reaction mixture was diluted with ethylacetate and hexane (1:1 ratio) and
washed with H.sub.2 O (20 mL), brine, dried over Na.sub.2 SO.sub.4. The
organic layer was concentrated in vacuo and isolated on a silica-gel
column using CHCl.sub.3 /EtOAc (50:1 ratio) as an eluting solvent to give
title compound (83% yield).
C. (S)-4-(2-Pyrrolidinylmethyl)amino!benzenecarboximidamide
To a solution of Part B nitrile (1.36 g, 4.52 mmol) in dry EtOH was bubbled
HCl gas at 0.degree. C. for 10 minutes. Sealed and stirred at room
temperature for 72 hrs. The reaction was concentrated in vacuo and was
redissolved in dry EtOH. NH.sub.3 gas was bubbled in the reaction until it
is basic and the reaction was stirred at room temperature for 24 hrs and
concentrated in vacuo to give title compound (quant. yield).
D.
S-(R*,S*)!-4-1-2-(1,1-Dimethylethoxy)carbonyl!amino!-1-oxo-3-phenyl
propyl!-2-pyrrolidinyl!methyl!amino!benzenecarboximidamide
To a solution of L-Boc-phenylalanine (1.45 g, 5.50 mmol) and HOBT (743 mg,
5.50 mmol) in 26 mL DMF was added WSC (1.05 g, 5.50 mmol) and Part C
compound (1.20 g, 5.50 mmol). After 5 minutes, NMM (605 .mu.l, 5.50 mmol)
was added to adjust the pH to 7.5-8.0. The reaction was stirred at room
temperature for 16 hrs and partitioned between EtOAc and H.sub.2 O. The
EtOAc layer was further washed with a sat. solution of NaHCO.sub.3, a sat.
solution of KHSO.sub.4, brine, dried over Na.sub.2 SO.sub.4 and
concentrated in vacuo. The crude product was subjected to a Prep. HPLC to
obtain title compound (42% yield).
E.
S-(R*,S*)!-4-1-(2-Amino-1-oxo-3-phenylpropyl)-2-pyrrolidinyl!methyl!am
ino!benzenecarboximidamide, trifluoroacetate (1:2) salt
To a solution of Part D compound (390 mg, 0.84 mmol) in 5 mL CH.sub.2
Cl.sub.2 was added TFA at 0.degree. C. Removed the ice bath and the
reaction stirred at room temperature for 2 hrs. The reaction mixture was
concentrated in vacuo and subjected to a Prep. HPLC to give title compound
(68% yield).
Elemental analysis C.sub.21 H.sub.27 H.sub.5 O.2.10 TFA.0.80 H.sub.2 O
Calc: C, 48.87; H, 5.00; N, 11.31; F, 19.33 Found: C, 49.13; H, 4.68; N,
10.89; F, 19.40.
EXAMPLE 17A
S-(R*,R*)!-4-(Aminoiminomethyl)amino!-N-1-3-hydroxy-2-(7-methoxy-2-n
apthalenyl)sulfonyl!amino!-1-oxopropyl!-2-pyrrolidinyl!methyl!butanamide,
trifluoroacetamide (1:1) salt
A.
S-(R*,R*!-1-2-(Azidomethyl)-1-pyrrolidinyl!-2-(1,1-dimethylethoxy)carb
onyl!amino!-3-(phenylmethoxy)-1-propanone
Example 1a Part A compound (2.13 g, 9.4 mmol) was dissolved in
dichloromethane (5 mL) and trifluoroacetic acid (7 mL) and stirred at RT 3
hours. The TFA and dichloromethane were removed by distillation under
reduced pressure and by coevaporation with toluene to give the amine as a
TFA salt. This and N-BOC-O-benzyl-L-serine (2.95 g, 10 mmol) were
dissolved in DMF (50 mL). HOBT (1.35 g, 10 mmol), WSC (1.91 g, 10 mmol)
and NMM (3.3 mL, 30 mmol) were added. The reaction was stirred for 8 h at
room temperature, diluted with ethyl acetate (50 mL) and satd. KHSO.sub.4
solution (30 mL). The layers were separated and the aqueous layer was
reextracted with ethyl acetate (50 mL). The combined organic layers were
washed with saturated sodium bicarbonate solution (30 mL)) and 10% lithium
chloride solution (30 mL). The ethyl acetate layer was dried over
magnesium sulfate and concentrated in vacuo to provide title compound
(3.87 g, 100%) as an oil which was used without further purification.
B.
S-(R*,R*)!-2-Amino-1-2-(azidomethyl)-1-pyrrolidinyl!-3-(phenylmethoxy)-1
-propanone
Part A BOC-azide (604 mg, 1.5 mmol) was dissolved in dichloromethane (1 mL)
and TFA (2 mL) and stirred at RT 3 hours. Solvents were removed by
distillation under reduced pressure and by coevaporation with toluene to
give title amine as the TFA salt which was used in the next step.
C.
S-(R*,R*)!-N-2-2-(Azidomethyl)-1-pyrrolidinyl!-2-oxo-1-(phenylmethoxy)
methyl!ethyl!-7-methoxy-2-naphthalenesulfonamide
The crude Part B TFA salt (1.5 mmol) was dissolved in dichloromethane (5
mL), cooled in an ice bath, and 7-methoxynapthene-2-sulfonyl chloride (384
mg, 1.5 mmol) was added, followed by dropwise addition of triethylamine (2
mL). The solution was stirred cold for 2 h before diluting with ethyl
acetate (30 mL). This solution was washed with 1N HCl solution (10 mL),
dried over magnesium sulfate and evaporated to provide crude title
sulfonamide. The crude product was chromatographed on silica gel and
eluted with 20 and 30% EtOAc in hexane to provide title sulfonamide as a
foam (536 mg, 68% overall in two steps).
D.
S-(R*,R*)-N-2-2-(Aminomethyl)-1-pyrrolidinyl!-2-oxo-1-(phenylmethoxy)m
ethyl!ethyl!-7-methoxy-2-naphthalenesulfonamide
The Part C sulfonamide (536 mg, 1 mmol) was dissolved in absolute ethanol
(15 mL), and treated with 10% palladium on carbon (105 mg). The flask was
equipped with a hydrogen filled balloon via a three way stopcock. Air
inside the flask was evacuated under reduced pressure and the flask was
then filled with hydrogen from the balloon. This process was repeated
three times. The mixture was stirred eight hours in the hydrogen
atmosphere. The catalyst was then removed by filtration through a pad of
MgSO.sub.4 and the pad was washed with ethyl acetate. The solvent was
removed in vacuo to give title compound as a light yellow oil (477 mg,
94%).
E.
S-(R*,R*)!-N-1-2-(7-Methoxy-2-naphthalenyl)sulfonyl!amino!-1-oxo-3-(
phenylmethoxy)propyl!-2-pyrrolidinyl!methyl!-4-(phenylmethoxy)carbonyl!am
ino!butanamide
Part D compound (477 mg, 0.96 mmol) and Z-4-aminobutyric acid (237 mg, 1.0
mmol) were dissolved in DMF (10 mL) at RT. HOBT (135 mg, 1.0 mmol), WSC
(191 mg, 1.0 mmol) and NMM (330 .mu.L) were added. The reaction was
stirred for 5 h, diluted with satd KHSO.sub.4 solution (15 mL) and water
(15 mL). The product was extracted into ethyl acetate (2.times.20 mL) and
washed with saturated NaHCO.sub.3 (15 mL) and 10% LiCl solution (15 mL),
dried over magnesium sulfate and concentrated in vacuo. The crude yellow
oil was chromatographed on silica gel, eluting with 50% EtOAc in hexane
followed by 1-3% MeOH in EtOAc to provide title compound as a foam (500
mg, 73%).
F.
S-(R*,R*)!-4-Amino-N-1-3-hydroxy-2-(7-methoxy-2-naphthalenyl)sulfony
l!amino!-1-oxopropyl!-2-pyrrolidinyl!methyl!butanamide
Part E compound (500 mg, 0.71 mmol) was dissolved in ethanol (60 mL) to
which acetyl chloride (2.5 mL) had been added and the mixture was treated
with 10% palladium on carbon (200 mg) and hydrogenated at 55 psi for 24 h.
The catalyst was removed by filtration and the residual catalyst was
washed with EtOH. The filtrate was concentrated in vacuo to obtain title
des-benzyl, des-CBZ product. This was used without purification.
G.
S-(R*,R*)!-4-(Aminoiminomethyl)amino!-N-1-3-hydroxy-2-(7-methoxy-2-
naphthalenyl)sulfonyl!amino!-1-oxopropyl!-2-pyrrolidinyl!methyl!butanamide,
trifluoroacetamide (1:1) salt
The Part F material (.about.0.71 mmol) was dissolved in ethanol (40 mL), to
which amidinesulfonic acid (124 mg, 1.0 mmol) and triethylamine (300
.mu.L, 2.1 mmol) were added. After 16 h the mixture was concentrated to
dryness. The crude material was purified by preparative HPLC (YMC S-10 ODS
50.times.500 mm column, eluting with 50% methanol in water, containing
0.1% TFA). Fractions containing clean title compound were combined and
lyophilized to provide a white solid (167 mg, 35%), Purity.gtoreq.98%.
.alpha.!.sub.D =-62.8.degree. (c=0.5, MeOH)
Analysis: Calcd for 1.15 TFA+0.80 H.sub.2 O: C, 46.44; H, 5.45; N, 12.36;
F, 9.64; S, 4.71. Found: C, 46.50; H, 5.34;, N, 12.23; F, 9.52; S, 4.85.
EXAMPLE 18A
R-(R*,S*)!-N-2-2-4-(Aminoiminomethyl)phenyl!amino!methyl!pyrrolidinyl
!-2-oxo-1-(phenylmethyl)ethyl!methanesulfonamide, trifluoroacetate (1:1)
salt
To a solution of Example 16a compound (155 mg, 0.25 mmol) in 7 mL dry
CH.sub.2 Cl.sub.2 and 3 mL THF was added Et.sub.3 N (139 ml, 1 mmol),
followed by methanesulfonyl chloride (46 ml, 0.58 mmol). The reaction was
stirred at room temperature for 2 hrs and concentrated in vacuo. The crude
product was subjected to a Prep. HPLC using 40 to 90% B for 45 minutes
gradient, A=90% H.sub.2 O/0.1% TFA; B=90% CH.sub.3 OH/0.1% TFA; column:
YMC S-10 ODS (c=18) to give title compound (69% yield).
Elemental analysis C.sub.22 H.sub.29 N.sub.5 O.sub.3 S.1.25 TFA.0.60
H.sub.2 O Calc: C, 49.30; H, 5.31; N, 11.73; F, 11.94 Found: C, 49.10; H,
5.18; N, 11.53; F, 12.10.
EXAMPLE 19A
1(S),2S!-1-(Aminoiminomethyl)-N-1-3-hydroxy-2-(2-naphthalenylsulfonyl)
amino!-1-oxopropyl!-2-pyrrolidinyl!methyl!-3-piperidinecarboxamide,
trifluoroacetate (1:1) salt
A.
S-(R*,R*)!-N-2-2-(Azidomethyl)-1-pyrrolidinyl!-2-oxo-1-(phenylmethoxy)
methyl!ethyl!-2-naphthalenesulfonamide
Example 17a Part A BOC-azide (3.85 mg, 9.55 mmol) was dissolved in
trifluoroacetic acid (TFA) (20 ml) and stirred at RT 1.5 hours. The TFA
was removed by distillation under reduced pressure and by coevaporation
with toluene. The residue was dissolved in dichloromethane (100 mL),
cooled in an ice bath, and triethylamine (4.0 mL, 28.7 mmol) and
2-naphthalene sulfonyl chloride (2.38 g, 10.5 mmol) were added. The
cooling bath was removed and the solution was stirred for 2 h. This
solution was washed with KHSO.sub.4 solution (25 mL.times.2) and
NaHCO.sub.3 solution (25 mL.times.2), dried over magnesium sulfate and
evaporated to provide crude title compound. The crude product was
chromatographed on silica gel to provide title sulfonamide as an oil
(2.597 g, 55%).
B.
S-(R*,R*)!-N-2-2-(Aminomethyl)-1-pyrrolidinyl!-2-oxo-1-(phenylmethoxy)
methyl!ethyl!-2-naphthalenesulfonamide
The Part A azide (2.5 g, 5 mmol) was dissolved in absolute ethanol (100 mL)
and treated with 10% palladium on carbon (300 mg). The flask was equipped
with a hydrogen filled balloon via a three way stopcock. Air inside the
flask was evacuated under reduced pressure and the flask was then filled
with hydrogen from the balloon. This process was repeated three times. The
mixture was stirred overnight in the hydrogen atmosphere. The catalyst was
then removed by filtration through Celite and the pad was washed with
ethanol. The solvent was removed from the filtrate in vacuo to give title
amine as a foam (2.236 g, 96%).
C. 1,3-Piperidinedicarboxylic acid, 1-(phenylmethyl)ester
Benzyl chloroformate (4 mL) was added dropwise to a cooled
(0.degree.-5.degree. C.), stirred solution of piperidine-3-carboxylic acid
(1.03 g, 8 mmol) in 1N aqueous NaOH solution (25 mL). During the twenty
minute addition, the pH was maintained between 8 and 9 by addition of 1N
NaOH solution. After addition was complete, the mixture was stirred cold
for an additional hour, maintaining the pH thoughout this period. The
reaction mixture was then extracted with ether (2.times.40 mL). The ether
extracts were discarded and the aqueous layer was acidified with 1N HCl
solution. The acidified aqueous layer was then extracted with
dichloromethane (2.times.40 mL). The combined extracts were dried over
magnesium sulfate and concentrated in vacuo to give title acid as a
viscous oil (1.73 g, 82%).
D.
3-(S)1-(S)-2-(Naphthalenylsulfonyl)amino!-1-oxo-3-(phenylmethoxy)pro
pyl!-2-pyrrolidinyl!methyl!amino!carbonyl!-1-piperidinecarboxylic acid,
phenylmethyl ester
Part B amine (820 mg, 1.75 mmol, and Part C acid (526 mg, 2.0 mmol) were
dissolved in DMF (20 mL) at RT. HOBT (270 mg, 2 mmol), 4-methyl morpholine
(1.5 mL) and WSC (400 mg, 2 mmol) were added. The reaction was stirred
overnight at room temperature. The mixture was then diluted with ethyl
acetate (50 mL), satd KHSO.sub.4 solution (10 mL) and water (20 mL). The
layers were separated and the aqueous layer was extracted with ethyl
acetate (50 mL). The combined organic layers were washed with satd
NaHCO.sub.3 solution (20 mL) and 10% lithium chloride solution (2.times.20
mL), dried over magnesium sulfate and concentrated in vacuo The crude
yellow foam was chromatographed on silica gel, eluting with 50% EtOAc in
hexane followed by 75% EtOAc in hexane and finally with EtOAc to provide
title sulfonamide as a foam (1.03 g, 82%).
E.
N-(S)1-(S)-3-Hydroxy-2-(naphthalenylsulfonyl)amino!-1-oxopropyl!-2-pyr
rolidinyl!methyl!-3-piperidinecarbamide
Part D compound (1.0 g, 1.4 mmol) was dissolved in ethanol (100 mL) to
which acetyl chloride (2.0 mL) had been added and the mixture was treated
with 10% palladium on carbon (300 mg) and hydrogenated at 55 psi for 44 h.
The catalyst was removed by filtration and the pad was washed with EtOH.
The filtrate was concentrated in vacuo to obtain crude amine hydrochloride
salt.
F.
1(S),2S!-1-(Aminoiminomethyl)-N-1-3-hydroxy-2-(2-naphthalenylsulfonyl
)amino!-1-oxopropyl!-2-pyrrolidinyl!methyl!-3-piperidenecarboxamide,
trifluoroacetate (1:1) salt
The crude Part E amine was dissolved in dimethylformamide (3 mL).
H-pyrazole-1-carboxamidine (226 mg, 1.54 mmol) and diisopropylethyl amine
(540 .mu.L, 3.1 mmol) were added. The mixture was stirred over a weekend
at room temperature. Ether (15 mL) was then added. Gummy material
precipitated. The ether was decanted and the precipitate was washed with
more ether. The gummy material was dissolved in methanol and taken to
dryness in vacuo. The remaining material was purified by preparative HPLC
(YMC S-10 ODS 50.times.500 mm column, eluting with 50% methanol in water,
containing 0.1% TFA). Fractions containing clean title compound were
combined and lyophilized to provide a white solid which was used to obtain
NMR spectra, recovered, dissolved in water (20 mL) and relyophilized to
give title compound (399 mg, 42%), Purity.gtoreq.98%.
.alpha.!.sub.D =-36.3.degree. (c=0.6, MeOH)
Analysis: Calcd for 1.25 TFA+0.1 H.sub.2 O: C, 48.94; H, 5.29; N, 12.45; F,
10.56; S, 4.75. Found: C, 49.03; H, 5.30;, N, 12.27; F, 10.47; S, 4.66.
EXAMPLE 20A
S-(R*,R*)!-1-(Aminoiminomethyl)-N-1-3-hydroxy-2-(2-naphthalenylsulfony
l)amino!-1-oxopropyl!-2-pyrrolidinyl!methyl!-4-piperidinecarboxamide,
trifluoroacetate (1:1) salt
A. 1,4-Piperidinedicarboxylic acid, 1-(phenylmethyl) ester
Benzyl chloroformate (4 mL) was added dropwise to a cooled
(0.degree.-5.degree. C.), stirred solution of piperidine-4-carboxylic acid
(1.03 g, 8 mmol) in 1N aqueous NaOH solution (25 mL). During the twenty
minute addition, the pH was maintained between 8 and 9 by addition of 1N
NaOH solution. After addition was complete, the mixture was stirred cold
for an additional hour, maintaining the pH thoughout this period. The
reaction mixture was then extracted with ether (2.times.40 mL). The ether
extracts were discarded and the aqueous layer was acidified with 1N HCl
solution. The acidified aqueous layer was then extracted with
dichloromethane (2.times.40 mL). The combined extracts were dried over
magnesium sulfate and concentrated in vacuo to give title acid as a
viscous oil (2.0 g, 95%).
B.
S-(R*,R,*)-4-1-2-(Naphthalenylsulfonyl)amino!-1-oxo-3-(phenylmethox
y)propyl!-2-pyrrolidinyl!methyl!amino!carbonyl!-1-piperidinecarboxylic
acid, phenylmethyl ester
Example 19a, Part B amine (820 mg, 1.75 mmol) and Part A acid (526 mg, 2.0
mmol) were dissolved in DMF (20 mL) at RT. HOBT (270 mg, 2 mmol), 4-methyl
morpholine (1.5 mL) and WSC (400 mg, 2 mmol) were added. The reaction was
stirred overnight at room temperature. The mixture was then diluted with
ethyl acetate (50 mL), satd KHSO.sub.4 solution. (10 mL) and water (20
mL). The layers were separated and the aqueous layer was extracted with
ethyl acetate (50 mL). The combined organic layers were washed with satd
NaHCO.sub.3 solution (20 mL) and 10% lithium chloride solution (2.times.20
mL), dried over magnesium sulfate and concentrated in vacuo. The crude
yellow foam was chromatographed on silica gel, eluting with 50% EtOAc in
hexane followed by 75% EtOAc in hexane and finally with EtOAc to provide
title sulfonamide as a foam (980 mg, 78%).
C.
S-(R*,R*)!-N-1-3-Hydroxy-2-(naphthalenylsulfonyl)amino!-1-oxopropyl!-
2-pyrrolidinyl!methyl!-4-piperidinecarbamide
Part B compound (970 mg, 1.36 mmol) was dissolved in ethanol (100 mL) to
which acetyl chloride (2.0 mL) had been added and the mixture was treated
with 10% palladium on carbon (400 mg) and hydrogenated at 55 psi for 30 h.
The catalyst was removed by filtration and the pad was washed with EtOH.
The filtrate was concentrated in vacuo to obtain crude title compound.
D.
S-(R*,R*)!-1-(Aminoiminomethyl)-N-1-3-hydroxy-2-(2-naphthalenylsulfon
yl)amino!-1-oxopropyl!-2-pyrrolidinyl!methyl!-4-piperidenecarboxamide,
trifluoroacetate (1:1) salt
The crude Part C amine was dissolved in dimethylformamide (3 mL).
H-pyrazole1-carboxamidine (226 mg, 1.54 mmol) and diisopropylethyl amine
(540 .mu.L, 3.1 mmol) were added. The mixture was stirred over a weekend
at room temperature. Ether (15 mL) was then added. Gummy material
precipitated. The ether was decanted and the precipitate was washed with
more ether. The gummy material was dissolved in methanol and taken to
dryness in vacuo. The remaining material was purified by preparative HPLC
(YMC S-10 ODS 50.times.500 mm column, eluting with 50% methanol in water,
containing 0.1% TFA). Fractions containing clean title compound were
combined and lyophilized to provide a white solid (33269-139-26) which was
used to obtain NMR spectra, recovered, dissolved in water (20 mL) and
relyophilized to give title compound (264 mg, 29%), Purity.gtoreq.98%.
.alpha.!.sub.D =-45.4.degree. (c=0.6, MeOH)
Analysis: Calcd for 1.25 TFA+0.1 H.sub.2 O: C, 48.94; H, 5.29; N, 12.45; F,
10.56; S, 4.75. Found: C, 49.12; H, 5.42;, N, 12.45; F, 10.62; S, 4.82.
EXAMPLE 21a
N-(S)-2-(S)-2-1-(Aminoiminomethyl)-3-piperidinyl!acetyl!amino!methyl!
-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
trifluoroacetate (1:1) salt
A.
S-(R*,R*)!-N-2-2-(Azidomethyl)-1-pyrrolidinyl!-2-oxo-1-(phenylmethoxy)
methyl!ethyl!-2-naphthalenesulfonamide
Example 17a, Part A BOC-Azide (3.85 mg, 9.55 mmol) was dissolved in
trifluoroacetic acid (TFA) (20 mL) and stirred at RT 1.5 hours. The TFA
was removed by distillation under reduced pressure and by coevaporation
with toluene. The residue was dissolved in dichloromethane (100 mL),
cooled in an ice bath, and triethylamine (4.0 mL, 28.7 mmol) and
2-naphthalene sulfonyl chloride (2.38 g, 10.5 mmol) were added. The
cooling bath was removed and the solution was stirred for 2 h. This
solution was washed with KHSO.sub.4 solution (25 mL.times.2) and
NaHCO.sub.3 solution (25 mL.times.2), dried over magnesium sulfate and
evaporated to provide crude title compound. The crude product was
chromatographed on silica gel and eluted with 30 and 50% EtOAc in hexane
to provide title sulfonamide as an oil (2.597 g, 55%).
B.
S-(R*,R*)!-N-2-2-(Aminomethyl)-1-pyrrolidinyl!-2-oxo-1-(phenylmethoxy)
methyl!ethyl!-2-naphthalenesulfonamide
The Part A azide (2.5 g, 5 mmol) was dissolved in absolute ethanol (100 mL)
and. treated with 10% palladium on carbon (300 mg). The flask was equipped
with a hydrogen filled balloon via a three way stopcock. Air inside the
flask was evacuated under reduced pressure and the flask was then filled
with hydrogen from the balloon. This process was repeated three times. The
mixture was stirred overnight in the hydrogen atmosphere. The catalyst was
then removed by filtration through Celite and the pad was washed with
ethanol. The solvent was removed from the filtrate in vacuo to give title
amine as a foam (2.236 g, 96%).
C. 1-(Phenylmethoxy)carbonyl!-3-piperidineacetic acid
Platinum oxide (300 mg, Alfa) was added to a solution of 3-pyridyl acetic
acid (3.3 g, 24.08 mmol) in water (120 mL) and conc. HCl (2.5 mL). The
solution was hydrogenated at up to 50 psi of H.sub.2 pressure for 6.5 hrs.
The catalyst was removed by filtration through a pad of Celite and the pad
was washed with more water. The stirred aqueous solution of the
3-piperidine acetic acid was cooled (0.degree.-5.degree. C.) and treated
with 5N aqueous NaOH solution (20 mL). Benzyl chloroformate (4 mL) was
added dropwise to the cooled, stirred solution. During the ten minute
addition, the pH was maintained between 10 and 12 by addition of NaOH
solution. After one hour the reaction mixture was extracted with ether
(2.times.100 mL). The ether extracts were discarded and the aqueous layer
was acidified with 2N HCl solution. The acidified aqueous layer was then
extracted with ethyl acetate (2.times.100 mL). The combined extracts were
dried over magnesium sulfate and concentrated in vacuo to give title
compound as an oil (7.0 g, contained EtOAc).
D.
N-(S)-2-Oxo-2-(S)-2-1-(phenylmethoxy)carbonyl!-3-piperidinyl!acetyl
!amino!methyl!-1-pyrrolidinyl!-1-(phenylmethoxy)methyl!ethyl!-2-naphthalen
esulfonamide
Part B amine (820 mg, 1.75 mmol), and Part C acid (554 mg, 2.0 mmol) were
dissolved in DMF (20 mL) at RT. HOBT (270 mg, 2 mmol), 4-methyl morpholine
(1.5 mL) and WSC (400 mg, 2 mmol) were added. The reaction was stirred 24
hours at room temperature. The mixture was then diluted with aqueous
KHSO.sub.4 solution and extracted with ethyl acetate (2.times.75 mL), The
combined organic layers were washed with satd. NaHCO.sub.3 solution (30
mL), dried over magnesium sulfate and concentrated in vacuo. The crude
yellow foam was chromatographed on silica gel, eluting with 50% EtOAc in
hexane, followed by 75% EtOAc in hexane, EtOAc and finally with 2% MeOH in
EtOAc to provide title compound as a colorless oil (900 mg, 71%).
E.
N-(S)1-(Hydroxymethyl)-2-oxo-2-(S)-2-1-(phenylmethoxy)carbonyl!-3-p
iperidinyl!acetyl!amino!methyl!-1-pyrrolidinyl!ethyl!-2-naphthalenesulfonam
ide, hydrochloride
Part D compound (900 mg, 1.24 mmol) was dissolved in ethanol (100 mL) to
which acetyl chloride (2.5 mL) had been added and the mixture was treated
with 10% palladium on carbon (250 mg) and hydrogenated at 55 psi for 24 h.
The catalyst was removed by filtration and the pad was washed with EtOH.
The filtrate was concentrated in vacuo to obtain crude title compound
which was used without purification.
F.
N-(S)-2-(S)-2-1-(Aminoiminomethyl)-3-piperidinyl!acetyl!amino!methyl
!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
trifluoroacetate (1:1) salt
The crude Part E amine was dissolved in dimethylformamide (3 mL).
H-pyrazole1-carboxamidine (226 mg, 1.54 mmol) and diisopropylethyl amine
(536 .mu.L, 3.1 mmol) were added. The mixture was stirred 3 days at room
temperature. Ether (15 mL) was then added. Gummy material precipitated.
The ether was decanted and the precipitate was washed with more ether. The
gummy material was dissolved in methanol and taken to dryness in vacuo.
The remaining material was purified by preparative HPLC (YMC S-10 ODS
50.times.500 mm column, eluting with 49% methanol in water, containing
0.1% TFA). Fractions containing clean title compound were combined and
lyophilized to provide a white solid (283 mg, 33%), Purity.gtoreq.98%.
.alpha.!.sub.D =-39.2.degree. (c=0.7, MeOH)
Analysis: Calcd for 1.2 TFA+0.4 H.sub.2 O: C, 49.53; H, 5.56; N, 12.20; F,
9.93; S, 4.66 Found: C, 49.54; H, 5.60; N, 12.16; F, 10.16; S, 4.79
EXAMPLE 22a
S-(R*,R*)!-N-2-2-1-(Aminoiminomethyl)-4-piperidinyl!acetyl!amino!met
hyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide
, trifluoroacetate (1:1) salt
A. 1-(Phenylmethoxy)carbonyl!-4-piperidineacetate acid
Platinum oxide (300 mg, Alfa) was added to a solution of pyridyl-4-acetic
acid, hydrochloride (3.3 g, 19 mmol) in water (120 mL). The solution was
hydrogenated at up to 50 psi of H.sub.2 pressure for 17 hrs. The catalyst
was removed by filtration through a pad of Celite and the pad was washed
with more water. The stirred aqueous solution of the piperidine-4-acetic
acid was cooled (0.degree.-5.degree. C.) and treated with 5N aqueous NaOH
solution (15 mL) to pH 12. Benzyl chloroformate (4 mL) was added dropwise
to the vigorously stirred cooled solution. After one hour (maintaining the
pH at 10 to 12), the reaction mixture was extracted with ether
(2.times.100 mL). The ether extracts were discarded and the aqueous layer
was acidified with 2N HCl solution (50 mL). The acidified aqueous layer
was then extracted with ethyl acetate (2.times.75 mL). The combined
extracts were dried over magnesium sulfate and concentrated in vacuo to
give title compound as a white solid (4.53 g, 86% yield).
B.
S-(R*,R*)!-N-2-Oxo-2-2-1-(phenylmethoxy)carbonyl!-4-piperidinyl!ac
etyl!amino!methyl!-1-pyrrolidinyl!-1-(phenylmethoxy)methyl!ethyl!-2-naphth
alenesulfonamide
Example 21a amine (820 mg, 1.75 mmol) and Part A acid (554 mg, 2.0 mmol)
were dissolved in DMF (20 mL) at RT. HOBT (270 mg, 2 mmol), 4-methyl
morpholine (1.5 mL) and WSC (400 mg, 2 mmol) were added. The reaction was
stirred 24 hours at room temperature. The mixture was then diluted with
aqueous KHSO.sub.4 solution (30 mL) and extracted with ethyl acetate
(2.times.75 mL). The combined organic layers were washed with satd.
NaHCO.sub.3 solution (50 mL), dried over magnesium sulfate and
concentrated in vacuo. The crude yellow foam was chromatographed on silica
gel, eluting with 50% EtOAc in hexane, followed by 70% EtOAc in hexane,
EtOAc and finally with 2% MeOH in EtOAc to provide title compound as a
colorless oil (1.0 g, 79%).
C.
S(R*,R*)!-N-1-(Hydroxymethyl)-2-oxo-2-2-1-(phenylmethoxy)carbonyl!
-4-piperidinyl!acetyl!amino!methyl!-1-pyrrolidinyl!ethyl!-2-naphthalenesulf
onamide, hydrochloride
Part B compound (1.0 g, 1.377 mmol) was dissolved in ethanol (100 mL) to
which acetyl chloride (2.5 mL) had been added and the mixture was treated
with 10% palladium on carbon (250 mg) and hydrogenated at 55 psi for 48 h.
The catalyst was removed by filtration and the pad was washed with EtOH.
The filtrate was concentrated in vacuo to obtain crude title compound
which was used without purification.
D.
S-(R*,R*)!-N-2-2-1-(Aminoiminomethyl)-4-piperidinyl!acetyl!amino!me
thyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamid
e, trifluoroacetate (1:1) salt
The crude Part C amine was dissolved in dimethylformamide (DMF) (3 mL).
H-pyrazole-1-carboxamidine (226 mg, 1.54 mmol) and diisopropylethyl amine
(536 .mu.L, 3.1 mmol) were added. The mixture was stirred 4 days at room
temperature, following the course of the reaction by TLC and HPLC during
this time. Ether (15 mL) was then added. Gummy material precipitated. The
ether was decanted and the precipitate was washed with more ether. The
gummy material was dissolved in methanol and taken to dryness in vacuo.
The remaining material was purified by preparative HPLC (YMC S-10 ODS
50.times.500 mm column, eluting with 47% methanol in water, containing
0.1% TFA). Fractions containing clean title compound were combined and
lyophilized to provide a white solid. (344 mg, 40%), Purity.gtoreq.98%.
.alpha.!.sub.D =-41.5.degree. (c=0.6, MeOH)
Analysis: Calcd for 1.2 TFA+1.0 H.sub.2 O: C, 48.76; H, 5.65; N, 12.01; F,
9.78; S, 4.58. Found: C, 48.76; H, 5.58; N, 12.06; F, 9.63; S, 4.73
EXAMPLE 23a
S-(R*,R*)!-N-2-2-5-(Aminoiminomethyl)amino!-1-oxopentyl!amino!methyl
!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
trifluoroacetate (1:1) salt
Following the procedure of Examples 1a and 2a except substituting in
Example 1a, Part C N-CBZ-5-aminopentanoic acid for N-CBZ-4-aminobutyric
acid, the title compound was obtained. The crude material was purified by
preparative HPLC (YMC S-10 ODS 50.times.500 mm column, eluting with 51%
methanol in water, containing 0.1% TFA). Fractions containing clean title
compound were combined and lyophilized to provide a white solid (199 mg,
46%), Purity.gtoreq.98%.
.alpha.!.sub.D =-43.5.degree. (c=0.6, MeOH)
Analysis: Calcd for 1.0TFA+0.70 H.sub.2 O: C, 48.40; H, 5.69; N, 13.02; F,
8.83; S, 4.97 Found: C, 48.42; H, 5.60; N, 12.84; F, 9.03; S, 4.79.
EXAMPLE 24a
S-(R*,R*)!-N-2-2-6-(Aminoiminomethyl)amino!-1-oxohexyl!amino!methyl!
-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
trifluoroacetate (1:1) salt
Following the procedure of Examples 1a and 2a except substituting
N-CBZ-6-aminohexanoic acid for N-CBZ-4-aminobutyric acid, the title
compund was prepared. The crude material was purified by preparative HPLC
to provide a white solid (160 mg), Purity.gtoreq.98%. .alpha.!.sub.D
=-44.3.degree. (c=0.5, MeOH)
Analysis: Calcd for 1.0 TFA+1.60 H.sub.2 O: C, 48.01; H, 6.00; N, 12.44; F,
8.44; S, 4.75 Found: C, 48.33; H, 5.70; N, 11.92; F, 8.18; S, 4.71.
EXAMPLE 25a
1S2R*(3R*)!!-N-2-2-1-(Aminoiminomethyl)-3-piperidinyl!carbonyl!amin
o!methyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfon
amide, trifluoroacetate (1:1) salt
A. (S)-3-Piperidinecarboxylic acid, ethyl ester, D-tartrate salt
Ref: Johnston, G. A. R. et al, J Neurochemistry, 1976, Vol 26,
pp.1029-1032.
Ethyl nipecotate (50 g, 318 mmol ) and D-tartaric acid (47.74 g, 318 mmol
)were dissolved in hot abs. ethanol (250 mL). A very small amount of
insoluble material was removed by filtration through a pad of Celite. The
filtrate gave crystalline material on cooling. This was harvested and
recrystallized eight times from absolute ethanol to give title compound as
a white solid (.about.27 g). m.p. 155.degree.-156.degree. C.,
.alpha.!.sub.365 =-200.7.degree. (c=2.0, 0.2% aqueous ammonium
molybdate).
B. (S)-1-(Phenylmethoxy)carbonyl!-3-piperidinecarboxylic acid
The Part A tartrate salt (1.0 g) was dissolved in water (5 mL) and a
solution of potassium carbonate was added to bring the pH to 9. The
solution was cooled in an ice water bath and ether (5 mL) was added.
Benzyl chloroformate (0.5 mL) was added dropwise to the well stirred
solution. During the addition, the pH was maintained between 8 and 9 by
addition of potassium carbonate solution. After addition was complete, the
mixture was stirred cold for an additional 1.5 hours, maintaining the pH
thoughout this period. The layers were separated and the aqueous was
reextracted with ether. The combined ether layers were dried over
magnesium sulfate and concentrated in vacuo. The ethyl ester obtained was
purified on silica gel, eluting with 25% ethyl acetate in hexane. The
purified material (930 mg) was dissolved in methanol (8 mL) and treated
with 1N NaOH solution (4 mL). After stirring at room temperature two
hours, the methanol was removed in vacuo. The aqueous solution was
acidified with 1N HCl and the acid was extracted into ethyl acetate
(2.times.10 mL). The combined extracts were dried over magnesium sulfate
and concentrated in vacuo to give title compound as a crystalline solid.
(773 mg). .alpha.!.sub.D =+49.9.degree. (c=1.4, MeOH). The optical purity
of this material was determined by coupling some of this material with
(S)-(-)-.alpha.-methyl-benzylamine. The material obtained was submitted to
the analytical HPLC group for determination of purity and was found to be
97.6%, (e.e.=95.2).
C.
1S2R*(3R*)!!-N-2-2-1-(Aminoiminomethyl)-3-piperidinyl!carbonyl!ami
no!methyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfo
namide, trifluoroacetate (1:1) salt
Following the procedure of Example 19a, the title compound was prepared
using the Example 19a, Part B amine and the nonracemic Part B acid to give
the crude product which was purified by preparative HPLC (360 mg, 46.4%),
Purity.gtoreq.98%.
.alpha.!.sub.D =-15.1.degree. (c=0.7, MeOH)
Analysis: Calcd for 1.20 TFA+0.4 H.sub.2 O: C, 48.78; H, 5.38; N, 12.46; F,
10.14; S, 4.75. Found: C, 48.71; H, 5.50;, N, 12.43; F, 10.12; S, 4.78.
EXAMPLE 26a
1S2R*(3S*)!!-N-2-2-1-(Aminoiminomethyl)-3-piperidinyl!carbonyl!amin
o!methyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfon
amide, trifluoroacetate (1:1) salt
A. (R)-3-Piperidinecarboxylic acid, ethyl ester, L-tartrate salt
Ref: Johnston, G. A. R. et al, J Neurochemistry, 1976, Vol. 26,
pp.1029-1032.
Ethyl nipecotate (50 g, 318 mmol ) and L-tartaric acid (47.74 g, 318 mmol
)were dissolved in hot abs. ethanol (200 mL). Crystalline material was
deposited on cooling. This was harvested and recrystallized 11 times from
absolute ethanol to give title compound as a white solid (.about.26 g).
m.p. 155.degree.-156.degree. C.
.alpha.!.sub.365 =+202.5.degree. (c=2.0, 0.2% aqueous ammonium molybdate).
B. (R)-1-(Phenylmethoxy)carbonyl!-3-piperidinecarboxylic acid
The Part A tartrate salt (1.0 g) was dissolved in water (5 mL) and a
solution of potassium carbonate was added to bring the pH to 9. The
solution was cooled in an ice water bath and ether (5 mL) was added.
Benzyl chloroformate (0.5 mL) was added dropwise to the well stirred
solution. During the addition, the pH was maintained between 8 and 9 by
addition of potassium carbonate solution. After addition was complete, the
mixture was stirred cold for an additional 1.5 hours, maintaining the pH
throughout this period. The layers were separated and the aqueous was
reextracted with ether. The combined ether layers were dried over
magnesium sulfate and concentrated in vacuo. The ethyl ester obtained was
purified on silica gel, eluting with 25% ethyl acetate in hexane. The
purified material (971 mg) was dissolved in methanol (8 mL) and treated
with 1N NaOH solution (4 mL). After stirring at room temperature two
hours, the methanol was removed in vacuo. The aqueous solution was
acidified with 1N HCl and the acid was extracted into ethyl acetate
(2.times.10 mL). The combined extracts were dried over magnesium sulfate
and concentrated in vacuo to give title compound as a crystalline solid.
(881 mg). .alpha.!.sub.D =-49.6.degree. (c=1.4, MeOH). The optical purity
of this material was determined by coupling some of this material with
(S)-(-)-.alpha.-methyl-benzylamine. The material obtained was submitted to
the analytical HPLC group for determination of purity and was found to be
97.3%, (e.e.=94.6).
C.
1S2R*(3S*)!!-N-2-2-1-(Aminoiminomethyl)-3-piperidinyl!carbonyl!ami
no!methyl-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfon
amide, trifluoroacetate (1:1) salt
The title compound was prepared employing the procedure of Example 19a
using Example 19a, Part B amine and nonracemic Parc B acid to give the
crude product which was purified by preparative HPLC (133 mg, 32%),
Purity.gtoreq.98%.
.alpha.!.sub.D =-60.920 (c=0.6, MeOH)
Analysis: Calcd for 1.05 TFA+1.1 H.sub.2 O: C, 48.57; H, 5.60; N, 12.54; F,
8.93; S, 4.78. Found: C, 48.52; H, 5.44; N, 12.31; F, 8.82; S, 4.82.
EXAMPLE 27a
S-(R*,R*)!-N-2-2-7-(Aminoiminomethyl)amino!-1-oxoheptyl!amino!methyl
!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
trifluoroacetate (1:1) salt
The title compound was prepared employing the procedure of Examples 1a and
2a except that N-CBZ-7-aminoheptanoic acid was employed in place of
N-CBZ-4-aminobutyric acid. The crude material was purified by preparative
HPLC to give (276 mg).
Purity.gtoreq.98%.
.alpha.!.sub.D =-42.3.degree. (c=0.6, MeOH)
Analysis: Calcd for 1.05 TFA+0.60 H.sub.2 O: C, 49.84; H, 5.99; N, 12.41;
F, 8.84; S, 4.73. Found: C, 49.77; H, 5.91; N, 12.34; F, 8.78; S, 4.93.
EXAMPLE 28a
S-(R*,R*)!-4-(Aminomethyl)-N-1-3-hydroxy-2-(2-naphthalenylsulfonyl)ami
no!-1-oxopropyl!-2-pyrrolidinyl!methyl!benzamide, trifluoroacetate (1:)
salt
##STR152##
To a stirred solution of 1-BOC-2-(methylazide)pyrrolidine (12.54 g, 55.5
mmol) (prepared employing a procedure similar to that described in Example
1 Part A) in 10 mL of dry dichloromethane at 0.degree. C. was added a
solution of 4N HCl in dioxane (25.0 mL, 100 mmol ). The solution was
stirred at room temperature for 2.5 h and concentrated in vacuo to give a
crude amine as an oil. To a stirred solution of this amine, N-BOC-L-serine
(11.4 g, 55.5 mmol ) and 1-hydroxybenzotriazole monohydrate (9.37 g, 55.5
mmol) in 240 mL of DMF was added in order N-methylmorpholine (18.3 mL, 167
mmol) and ethyl 3-(3-dimethyl amino)propyl carbodiimide hydrochloride
(10.6 g, 55.5 mmol). The reaction solution was stirred at room temperature
for 19 h and concentrated under pump vacuum at 45.degree. C. The residue
was dissolved in 1 L of EtOAc and washed with 5% KHSO.sub.4 solution
(3.times.0.5 L), saturated NaHCO.sub.3 solution (2.times.0.5 L) and brine
(1.times.0.5 L). The EtOAc layer was dried (MgSO.sub.4), filtered and
concentrated in vacuo to give 14.4 g (83%) of title A(1) amide.
##STR153##
To a stirred solution of Part A(1) amide (14.3 g, 45.9 mmol) in 20 mL of
dry dichloromethane at 0.degree. C. was added a solution of 4N HCl in
dioxane (40.0 mL, 160 mmol). The solution was stirred at room temperature
for 2 h and concentrated in vacuo to give a crude amine as an oil. To a
stirred solution of this amine and triethylamine (15.4 mL, 110 mmol) in
100 mL of dry dichloromethane at 0.degree. C. was added dropwise a
solution of 2-naphthalenesulfonyl chloride (10.9 g, 48.2 mmol) in 60 mL of
dry dichloromethane over 40 min. The reaction was stirred at 0.degree. C.
for 1 h and at room temperature for 2 h. The solution was diluted with 1 L
of dichloromethane and washed with 1N HCl solution (3.times.0.5 L),
saturated NaHCO.sub.3 solution (2.times.0.5 L) and brine (1.times.0.5 L).
The dichloromethane layer was dried (MgSO.sub.4), filtered and
concentrated in vacuo. This was triturated in EtOAc-hexane to give 11.5 g
of title A(2) azide. The triturant was concentrated in vacuo and
chromatographed on silica gel to give 12.9 g of title A(2) azide.
##STR154##
To a stirred solution of Part A(2) azide (11.2 g, 27.8 mmol) in 300 mL of
EtOH and 600 mL of methanol was added 10% Pd/C (2.24 g). The atmosphere
was replaced with hydrogen and the reaction mixture stirred at room
temperature for 17 h. The catalyst was filtered off through a 4 .mu.M
polycarbonate film and rinsed with methanol (3.times.100 mL). The filtrate
was concentrated in vacuo to give 9.9 g (94%) of crude title A amine.
##STR155##
To a solution of 4-(aminomethyl)benzoic acid (10.33 g, 68.3 mmol) in
ethanol (137 mL) and water (68 mL) were added aqueous NaOH (10M, 7.6 mL,
76.0 mmol) and di-tert-butyl dicarbonate (16.48 g, 75.5 mmol). After 24 h,
the reaction mixture was concentrated in vacuo. The residue was dissolved
in EtOAc and 10% aqueous KHSO.sub.4, the layers separated, and the aqueous
layer was extracted with EtOAc. The organic layers were combined, dried
over Na.sub.2 SO.sub.4, filtered and concentrated in vacuo to give title
compound (16.88 g, 98%) as a colorless solid.
##STR156##
To a solution of Part B acid (0.23 g, 0.93 mmol) and 1-hydroxybenzotriazole
hydrate (0.13 g, 0.94 mmol) in DMF (2.8 mL) at 0.degree. C. was added
ethyl-3-(3-dimethylamino)propyl carbodiimide.HCl (0.17 g, 0.91 mmol).
After 0.5 h, Part A amine (0.31 g, 0.83 mmol) in DMF (1.7 mL) was added,
followed by 4-methylmorpholine (0.12 mL, 1.1 mmol). The reaction mixture
was stirred for 15.5 h while allowing the reaction to warm to room
temperature. The reaction mixture was poured into water:brine (1:1) and
extracted with EtOAc, the organic layer washed with 0.25M aqueous
KHSO.sub.4, water, saturated aqueous NaHCO.sub.3, water and brine. The
organic layer was dried over Na.sub.2 SO.sub.4, filtered and concentrated
in vacuo to give title compound (0.50 g, 98%) as a colorless foam.
D.
S-(R*,R*)!-4-(Aminomethyl)-N-1-3-hydroxy-2-(2-naphthalenylsulfonyl)ami
no!-1-oxopropyl!-2-pyrrolidinyl!methyl!benzamide, trifluoroacetate (1:1)
salt
##STR157##
To a solution of Part C compound (0.44 g, 0.72 mmol) in CH.sub.2 Cl.sub.2
at 0.degree. C. was added trifluoroacetic acid (0.9 mL). After 1.5 h, the
reaction mixture was concentrated in vacuo and purified by preparative
HPLC. The appropriate fractions were combined, concentrated in vacuo,
dissolved in H.sub.2 O and lyophilized to yield title compound (0.14 g,
32%) as a colorless solid:
.alpha.!.sub.D =-38.6.degree. (c 1.03, MeOH)
MS (M+H).sup.+ =511.sup.+
Anal. Calc'd for C.sub.26 H.sub.30 N.sub.4 O.sub.5 S.1.1 TFA.1.33 H.sub.2
O: C, 51.32; H, 5.16; N, 8.49; S, 4.86; F, 9.50 Found: C, 51.32; H, 4.92;
N, 8.42; S, 5.03; F, 9.82
EXAMPLE 29a
##STR158##
Example 28a Part A amine (1.93 g, 5.15 mmol), 4-cyanobenzoic acid (757 mg,
5.15 mmol) and 1-hydroxybenzotriazole (869 mg, 6.44 mmol) were dissolved
in DMF (30 mL) at RT. 4-Methylmorpholine (1.1 mL, 10 mmol) was added
dropwise followed by WSC (1.03 g, 5.15 mmol). The reaction was stirred 20
hours at room temperature. The mixture was then diluted with ethyl acetate
(75 mL) and KHSO.sub.4 solution (60 mL), the layers were separated and the
aqueous layer was reextracted with ethyl acetate (75 ML). The combined
organic layers was washed with saturated. NaHCO.sub.3 solution (50 mL) and
10% lithium chloride solution (2.times.25 mL), dried over magnesium
sulfate and concentrated in vacuo. The crude oil was chromatographed on
silica gel, eluting with 50% EtOAc in hexane followed by 75% EtOAc in
hexane, EtOAc and finally 3% MeOH in EtOAc to provide title compound as a
foam (2.01 g, 78%).
##STR159##
Sodium pellets (.about.100 mg) were added to methanol (35 mL) and stirred
until all the sodium had been consumed. Part A compound (759 mg, 1.5 mmol)
was added and the mixture was stirred at room temperature. After three
hours, additional sodium (.about.50 mg) was added and the mixture was
stirred an additional 2 hours. After this time an equilibrium mixture of
product and starting material had been obtained. Ammonium chloride (4 g)
was added and the mixture was left stirring overnight at room temperature.
The reaction mixture was then acidified with 1N HCl. Insoluble material
was removed by filtration through Celite and the pad was washed with more
methanol. The filtrate was freed of solvent in vacuo. The remaining
material was purified by preparative HPLC (YMC S-15 ODS 50.times.500 mm
column, eluting with 49% methanol in water, containing 0.1% TFA).
Fractions containing clean title compound were combined and lyophilized to
provide a white solid which was used to obtain NMR spectra, recovered,
dissolved in water (40 mL) and relyophilized to give title compound (283
mg, 28%).
.alpha.!.sub.D =-37.6.degree. (c=0.5, MeOH)
Analysis: Calcd for 1.08 TFA+2.1 H.sub.2 O: C, 49.41; H, 5.05; N, 10.23; F,
8.99; S, 4.68 Found: C, 49.02; H, 4.57; N, 10.13; F, 8.70; S, 5.12.
EXAMPLE 30a
##STR160##
Ref: J Org. Chem, 53, 3513-3521, 1988.
Potassium permanganante (29.5 g, 187 mmol) was dissolved in water (450 mL)
and 3,5-lutidine (10 g, 93 mmol) was added. After stirring for 1 hour the
temperature was 40.degree. C. and the mixture was heated in an oil bath
maintained at 40.degree.-50.degree. C. overnight while the stirring was
continued. After cooling the mixture was filtered through Celite. The
colorless filtrate was concentrated to .about.50 mL and then acidified
with HCl. White solid precipitated and was harvested by filtration and
washed with more water. Mass Spec indicated this was a mixture of the
desired monoacid (A) and diacid (B) and was used in the next reaction.
(5.2 g).
##STR161##
The mixture of Parts A and B acids was suspended in absolute ethanol (100
mL) in a Parr bottle. Acetyl chloride (5 mL) and platinum oxide (165 mg)
were added and the mixture was hydrogenated at up to 55 psi for 20 hours.
The catalyst was removed by filtration and the pad was washed with
ethanol. The filtrate was taken to dryness in vacuo to give title
compound. Mass spec indicated this was a mixture of the ethyl ester (D)
and diester (D).
##STR162##
The mixture of Parts C and D ethyl esters was dissolved in water (50 mL)
and a solution of potassium carbonate was added to bring the pH to 8.5.
The solution was cooled in an ice water bath and ether (50 mL) was added.
Benzyl chloroformate (8 mL) was added dropwise to the well stirred
solution. During the addition, the pH was maintained between 8 and 9 by
addition of potassium carbonate solution. After addition was complete, the
mixture was stirred cold for an additional hour, maintaining the pH
thoughout this period. The layers were separated and the aqueous was
reextracted with ether. The combined ether layers were dried over
magnesium sulfate and concentrated in vacuo. The material was
chromatographed on silica gel, eluting with 10-20% ethyl acetate in
hexane. The cis and trans isomers were separated. An NMR study of the
acids indicated that the faster moving isomer (3.665 g) was the cis
material.
##STR163##
A portion of the purified Part E cis compound (1.525 g, 5 mmol)) was
dissolved in methanol (16 mL) and treated with 1N NaOH solution (10 mL).
After stirring at room temperature two hours, the methanol was removed in
vacuo. The aqueous solution was acidified with 1N HCl and the acid was
extracted into ethyl acetate (3.times.10 mL). The combined extracts were
dried over magnesium sulfate and concentrated in vacuo to give title
compound as a viscous oil (1.39 g, 100%).
##STR164##
2-Azido-N-BOC pyrrolidine (2.13 g, 9.4 mmol) was dissolved in
dichloromethane (5 mL) and trifluoroacetic acid (7 mL) and stirred at RT 3
hours. The TFA and dichloromethane were removed by distillation under
reduced pressure and by coevaporation with toluene to give the amine as a
TFA salt. This and N-BOC-O-benzyl-L-serine (2.95 g, 10 mmol) were
dissolved in DMF (50 mL). HOBT (1.35 g, 10 mmol), WSC (1.91 g, 10 mmol)
and NMM (3.3 mL, 30 mmol) were added. The reaction was stirred for 8 h at
room temperature, diluted with ethyl acetate (50 mL) and saturated
KHSO.sub.4 solution (30 mL). The layers were separated and the aqueous
layer was reextracted with ethyl acetate (50 mL). The combined organic
layers were washed with saturated sodium bicarbonate solution (30 mL) and
10% lithium chloride solution (30 mL). The ethyl acetate layer was dried
over magnesium sulfate and concentrated in vacuo to provide title G(1)
compound (3.87 g, 100%) as an oil which was used without further
purification.
##STR165##
Part G(1) BOC-azide (3.85 mg, 9.55 mmol) was dissolved in TFA (20 mL) and
stirred at RT 1.5 hours. The TFA was removed by distillation under reduced
pressure and by coevaporation with toluene. The residue was dissolved in
dichloromethane (100 mL), cooled in an ice bath, and triethylamine (4.0
mL, 28.7 mmol) and 2-naphthalene sulfonyl chloride (2.38 g, 10.5 mmol)
were added. The cooling bath was removed and the solution was stirred for
2 h. This solution was washed with KHSO.sub.4 solution (25 mL.times.2) and
NaHCO.sub.3 solution (25 mL.times.2), dried over magnesium sulfate and
evaporated to provide crude title sulfonamide. The crude title sulfonamide
product was chromatographed on silica gel and eluted with 30% and 50%
EtOAc in hexane to provide A(1) sulfonamide as an oil (2.597 g, 55%).
##STR166##
The Part G(2) azide (2.5 g, 5 mmol) was dissolved in absolute ethanol (100
mL) and treated with 10% palladium on carbon (300 mg). The flask was
equipped with a hydrogen filled balloon via a three way stopcock. Air
inside the flask was evacuated under reduced pressure and the flask was
then filled with hydrogen from the balloon. This process was repeated
three times. The mixture was stirred overnight in the hydrogen atmosphere.
The catalyst was then removed by filtration through Celite and the pad was
washed with ethanol. The solvent was removed from the filtrate in vacuo to
give title amine as a foam (2.236 g, 96%).
##STR167##
Part G amine (467 mg, 1.0 mmol) and Part F acid (305 mg, 1.1 mmol) were
dissolved in DMF (2 mL) at RT. HOBT (148 mg, 1.1 mmol), 4-methyl
morpholine (242 .mu.L, 2.2 mmol) and WSC (210 mg, 1.1 mmol) were added.
The reaction was stirred 20 hours at room temperature. The mixture was
then diluted with ethyl acetate (50 mL) and KHSO.sub.4 solution (20 mL).
The layers were separated. The organic layer was washed with saturated
NaHCO.sub.3 solution (20 mL) and 10% lithium chloride solution (2.times.20
mL), dried over magnesium sulfate and concentrated in vacuo. The crude oil
was chromatographed on silica gel, eluting with 50% EtOAc in hexane
followed by EtOAc to provide title compound (584 mg, 80%).
##STR168##
Part H compound (578 mg, 0.79 mmol) was dissolved in ethanol (100 mL) to
which acetyl chloride (2.0 mL) had been added and the mixture was treated
with 10% palladium on carbon (300 mg) and hydrogenated at 55 psi for 3
days. The catalyst was removed by filtration and the pad was washed with
EtOH. The filtrate was concentrated in vacuo to obtain crude title
compound as a foam (350 mg) which was used without purification.
##STR169##
The crude Part J amine (.about.0.79 mmol) was dissolved in
dimethylformamide (1.6 mL). H-pyrazole-1-carboxamidine (131 mg, 0.89 mmol)
and diisopropylethyl amine (310 .mu.L) were added. The mixture was stirred
24 hours at room temperature. Ether (10 mL) was then added. Gummy material
precipitated. The ether was decanted and the precipitate was washed with
more ether. The gummy material was dissolved in methanol and taken to
dryness in vacuo. The remaining material was purified by preparative HPLC
(YMC S-15 ODS 50.times.500 mm column, eluting with 54% methanol in water,
containing 0.1% TFA). The two cis isomers were separated. The fractions
that were clean by analytical HPLC were combined, concentrated to a small
volume in vacuo and lyophilized. These samples were used to obtain NMR
spectra, recovered, dissolved in water and relyophilized to give title
compound, isomer A (from Part H compound) and Example 48 compound, Isomer
B (124 mg, 22% from Part H compound). Also obtained was a small amount of
mixed fractions (54 mg, .about.10%). Title compound, Isomer A:
.alpha.!.sub.D =-49.0.degree. (c=0.4, MeOH)
Analysis: Calcd for 1.15 TFA+1.3 H.sub.2 O: C, 48.61; H, 5.73; N, 12.02; F,
9.37; S, 4.59. Found: C, 48.59; H, 5.62; N, 11.92; F, 9.35; S, 4.74.
EXAMPLE 31a
##STR170##
To a stirred solution of 1-BOC-2-(methylazide)pyrrolidine (3.00 g, 13.3
mmol) in 80 mL of methanol under argon was added 20% Pd(OH).sub.2 /C (0.60
g, 20% based on the weight of the azide compound). The atmosphere was
replaced with hydrogen by several vacuum-fill cycles. The reaction mixture
was stirred at room temperature for 19 h. The catalyst was filtered off
through a 4 .mu.M polycarbonate film and rinsed with methanol (4.times.30
mL). The filtrate was concentrated in vacuo to give 2.65 g of an
intermediate amine in a quantitative yield. To a stirred solution of this
amine, 1-hydroxybenzotriazole monohydrate (2.62 g, 15.5 mmol) and
N-carbobenzyloxy-3-piperidine carboxylic acid (4.08 g, 15.5 mmol) in 85 mL
of DMF was added in order N-methylmorpholine (8.51 mL, 77.5 mmol) and
ethyl 3-(3dimethylamino)propyl carbodiimide hydrochloride (5.94 g, 31.0
mmol). The reaction solution was stirred at room temperature for 18 h and
concentrated under pump vacuum at 50.degree. C. The residue was dissolved
in 500 mL of EtOAC and washed with 1N HCl solution (3.times.200 mL),
saturated NaHCO.sub.3 solution (2.times.200 mL) and brine (1.times.200
mL). The EtOAc layer was dried (MgSO.sub.4), filtered and concentrated in
vacuo. Purification was effected by flash chromatography on silica gel to
give 3.33 g (56%) of title carbamate.
##STR171##
To a stirred solution of Part A carbamate (3.20 g, 7.19 mmol) in 10 mL of
dry dichloromethane at 0.degree. C. was added a 4N HCl solution in dioxane
(15.0 mL, 60.0 mmol). The solution was stirred at room temperature for 3 h
and concentrated in vacuo. The residue was dissolved in 100 mL of
dichloromethane and 100 mL of methanol. The solution was concentrated in
vacuo to give an intermediate amine. To a stirred solution of this amine
(1.10 g, 2.88 mmol), 1-hydroxybenzotriazole monohydrate (0.48 g, 2.88
mmol) and N-Boc-O-benzylhomoserine (0.89 g, 2.88 mmol) in 20 mL of DMF was
added in order N-methylmorpholine (1.04 mL, 9.51 mmol) and ethyl
3-(3-dimethylamino)propyl carbodiimide hydrochloride (0.55 g, 2.88 mmol).
The reaction solution was stirred at room temperature for 18 h and
concentrated under pump vacuum at 45.degree. C. The residue was dissolved
in 300 mL of EtOAc and washed with 1N HCl solution (3.times.100 mL),
saturated NaHCO.sub.3 solution (2.times.100 mL) and brine (1.times.100
mL). The organic layer was dried (MgSO.sub.4), filtered and concentrated
in vacuo and chromatographed on silica gel to give 1.28 g (70%) of title
amide.
##STR172##
To a stirred solution of Part B amide (804 mg, 1.27 mmol) in 5.0 mL of dry
dichloromethane at 0.degree. C. was added a 4N HCl solution in dioxane
(10.0 mL, 40.0 mmol). The reaction solution was stirred at room
temperature for 3 h and concentrated in vacuo to give a crude amine. To a
stirred solution. of this amine and triethyl amine (0.39 mL, 2.78 mmol) in
20 mL of dry dichloromethane under argon was added 2-naphthalenesulfonyl
chloride (301 mg, 1.33 mmol). The reaction solution was stirred at room
temperature for 3.5 h and diluted with 200 mL of EtOAc. The solution was
washed with 1N HCl solution (3.times.60 mL), saturated NaHCO.sub.3
solution (2.times.60 mL) and brine (1.times.60 mL). The organic layer was
dried (MgSO.sub.4), filtered, concentrated in vacuo and was
chromatographed on silica gel to give 0.79 g (86%) of title carbamate.
##STR173##
To a stirred solution of Part C carbamate 0.71 g, 0.98 mmol) in 10 mL of
methanol under argon was added 20% Pd(OH).sub.2 /C (142 mg, 20% based on
the weight of Part C compound). The atmosphere was replaced with hydrogen
by several vacuum-fill cycles. The reaction mixture was stirred at room
temperature for 20 h. The catalyst was filtered off through a 4 .mu.M
polycarbonate film and rinsed with methanol (5.times.30 mL). The filtrate
was concentrated in vacuo to give 0.58 g of the intermediate amine. To a
stirred solution of this amine (124 mg, 0.21 mmol) and DIEA (73.0 mL, 0.42
mmol) in 0.25 mL of DMF was added guanopyrazole hydrochloride (37.0 mg,
0.25 mmol). The reaction solution was stirred at room temperature for 24 h
at which time another batch of guanopyrazole hydrochloride (6.20 mg, 0.04
mmol) was added. The solution was stirred at room temperature for 22 h.
The reaction solution was then diluted with 20 mL of ether and treated
with 0.15 mL of a solution of 4N HCl in dioxane. The solvent was decanted
and the precipitate was purified by preparative HPLC. The fractions were
concentrated in vacuo and lyophilized to give 118 mg (73%) of title
guanidine.
Analysis: calc'd for 1.00 TFA+1.00 H.sub.2 O: C, 54.82; H, 5.91; N, 10.96;
F, 7.43; S, 4.18 Found: C, 54.87; H, 5.83; N, 10.58; F, 7.82; S, 4.25.
EXAMPLE 32a
##STR174##
To a stirred solution of (.+-.) ethyl nipecotate (5.00 g, 31.8 mmol) and
triethyl amine (6.64 mL, 47.7 mmol) in 100 mL of dichloromethane at
0.degree. C. was added portionwise a solution of di-t-butyl dicarbonate
(7.64 g, 35.0 mmol) in 100 mL of dichloromethane over 5 min. The reaction
solution was stirred at room temperature for 25 h and then diluted with
600 mL of EtOAc. The solution was washed with 1N HCl solution (3.times.200
mL), saturated NaHCO.sub.3 solution (2.times.200 mL) and brine
(1.times.200 mL). The organic layer was dried (MgSO.sub.4), filtered and
concentrated in vacuo. This was chromatographed on silica gel to give 6.44
g (79%) of title N-Boc nipecotate.
##STR175##
To a stirred solution of Part A nipecotate (1.00 g, 3.91 mmol) in 40 mL of
dry THF under argon at -78.degree. C. was added a 1M solution of sodium
hexamethyldisilazide (4.30 mL, 4.30 mmol) over 5 min. The solution was
stirred at -78.degree. C. for 40 min at which time a solution of diphenyl
diselenide (1.28 g, 4.10 mmol) in 6.0 mL of THF was added dropwise over 10
min. This reaction solution was stirred at -78.degree. C. for 1 h and
quenched dropwise with 15 mL of saturated NaHCO.sub.3 solution. The
resulting mixture was stirred vigorously without cooling for 5 min. The
mixture was concentrated in vacuo. The residue was diluted with 300 mL of
EtOAc and washed with 5% KHSO.sub.4 solution (3.times.150 mL), saturated
NaHCO.sub.3 solution (2.times.150 mL) and brine (1.times.150 mL). The
organic layer was dried (MgSO.sub.4), filtered, concentrated in vacuo; and
purified on silica gel to give 1.29 g (80%) of Part B ester.
##STR176##
To a stirred solution of Part B ester (1.06 g, 2.57 mmol) in 65 mL of
methanol and 20 mL of water was added 1N NaOH solution (7.69 mL, 7.69
mmol). The reaction solution was stirred at room temperature for 10 days.
The solution was concentrated in vacuo. The remaining aqueous solution was
acidified to pH 2 by the addition of 1N HCl solution. The solution was
extracted with EtOAc (4.times.60 mL). The combined EtOAc extracts were
dried (MgSO.sub.4), filtered, concentrated in vacuo, and chromatographed
on silica gel to give 350 mg (36%) of title acid and some recovered
starting Part B ester.
##STR177##
To a stirred solution of Part C acid (263 mg, 0.68 mmol), Example 30a Part
G amine (283 mg, 0.68 mmol) and 1-hydroxybenzotriazole monohydrate (115
mg, 0.68 mmol) in 4.4 mL of DMF was added in order N-methylmorpholine
(0.23 mL, 2.05 mmol) and ethyl 3-(3-dimethylamino)propyl carbodiimine
hydrochloride (131 mg, 0.68 mmol). The reaction solution was stirred at
room temperature for 16 h and concentrated under pump vacuum at 45.degree.
C. The residue was dissolved in 200 mL of EtOAc and washed with 1N HCl
solution (2.times.60 mL), saturated NaHCO.sub.3 solution (2.times.60 mL)
and brine (1.times.60 mL). The EtOAc layer was dried (MgSO.sub.4),
filtered and concentrated in vacuo, and was chromatographed on silica gel
to give 417 mg (82%) of title carbamate.
##STR178##
To a stirred solution of Part D carbamate (415 mg, 0.56 mmol) in 3.0 mL of
dry dichloromethane was added TFA (9.00 mL, 117 mmol). The reaction
solution was stirred at room temperature for 1 h and concentrated in vacuo
to give title amine.TFA (384 mg, 91%).
##STR179##
To a stirred solution of Part E amine.TFA (381 mg, 0.50 mmol) and DIEA
(0.26 mL, 1.51 mmol) in 0.5 mL of DMF was added guanopyrazole
hydrochloride (88.0 mg, 0.60 mmol). The reaction solution was stirred at
room temperature for 24 h at which time another of batch of guanopyrazole
hydrochloride (14.7 mg, 0.10 mmol) was added. The reaction solution was
stirred at room temperature for 24 h and stood at room temperature for 21
h. The solution was diluted with 20 mL of ether. To this mixture was added
0.3 mL of a solution of 4N HCl in dioxane. The solution was decanted and
the precipitated residue was purified by preparative HPLC. The fractions
were concentrated in vacuo and lyophilized to give 236 mg (59%) of title
selenide.
##STR180##
To a stirred solution of Part F selenide (190 mg, 0.24 mmol) in 9 mL of THF
was added 1.6 mL of 30% H.sub.2 O.sub.2 solution. The reaction solution
was stirred at room temperature for 30 min, concentrated in vacuo and
purified by preparative HPLC to give 95 mg (62%) of a 1:1 mixture of title
G and H compounds.
MS: (M+H).sup.+ =529.
EXAMPLE 33a
##STR181##
A solution of 1-BOC-2-(methylazido)pyrrolidine (1.36 g, 6.0 mmol) in
dichloromethane (5 mL) and trifluoroacetic acid (4 mL) was stirred at room
temperature for two hours. The reaction mixture was concentrated under
reduced pressure and toluene was added and removed at reduced pressure.
This process was repeated two more times. The crude amine, N-BOC-D-alanine
(1.14 g, 6 mmol) and 1-hydroxybenzotriazole (1.01 g, 7.5 mmol) were
dissolved in DMF (30 mL) and treated with 4-methyl morpholine (1.32 mL, 12
mmol) followed by WSC (1.2 g, 6.0 mmol). The reaction mixture was stirred
overnight at room temperature. The mixture was diluted with ethyl acetate
(70 mL) and washed with saturated KHSO.sub.4 solution (20 mL), NaHCO.sub.3
solution (25 mL) and 10% lithium chloride solution (2.times.15 mL), dried
over magnesium sulfate and concentrated in vacuo to give title compound as
an oil (1.74 g, 98%) which was used without purification.
##STR182##
A solution of Part A azide (1.74 g, 5.85 mmol) in dichloromethane (5 mL)
and trifluoroacetic acid (4 mL) was stirred at room temperature for two
hours. The reaction mixture was concentrated under reduced pressure and
toluene was added and removed at reduced pressure. This process was
repeated two more times. A solution of the crude des-BOC-amine and
.alpha.-toluenesulfonyl chloride in dichloromethane (30 mL) was cooled to
0.degree.-5.degree. C. and triethylamine (4.2 mL, 30 mmol) was added
dropwise. The mixture was stirred cold for one hour and then diluted with
1N HCl solution (50 mL). The dichloromethane layer was separated, dried
over magnesium sulfate and concentrated in vacuo. The crude oil was
chromatographed on silica gel, eluting with 50% and 75% ethyl acetate in
hexanes to give the title benzylsulfonamide as a white foam (1.55 g, 75%).
##STR183##
The Part B azide (1.55 g, 4.42 mmol) was dissolved in absolute ethanol (60
mL) and treated with 10% palladium on carbon (310 mg). The reaction flask
was equipped with a hydrogen filled balloon via a three-way stopcock. Air
inside the flask was evacuated under reduced pressure and replaced with
hydrogen from the balloon. This operation was repeated (3.times.).
Hydrogenolysis was continued for twenty hours. The balloon was removed and
the catalyst was removed by filtration. The pad was washed with more
ethanol. The filtrate was concentrated in vacuo to obtain title amine
(1.33 mg, 93%) as a foam which was used without purification.
##STR184##
Part C amine (488 mg, 1.5 mmol) and N-carbobenzyloxy-3(S)-piperidine
carboxylic acid (395 mg, 1.5 mmol) were dissolved in DMF (15 mL) at RT.
HOBT (270 mg, 2.0 mmol), 4-methyl morpholine (550 .mu.L, 5 mmol) and WSC
(300 mg, 1.5 mmol) were added. The reaction was stirred 20 hours at room
temperature. The mixture was then diluted with ethyl acetate (50 mL) and
washed with KHSO.sub.4 solution (30 mL), saturated NaHCO.sub.3 solution
(25 mL) and 10% lithium chloride solution (3.times.15 mL), dried over
magnesium sulfate and concentrated in vacuo to give title compound (730
mg, 85%) as a white foam.
##STR185##
Part D compound (730 mg, 1.28 mmol) was dissolved in ethanol (100 mL) and
treated with Pearlman's catalyst (300 mg). The reaction flask was
connected to a hydrogen filled balloon via a three-way stopcock. Air
inside the flask was evacuated under reduced pressure and replaced with
hydrogen from the balloon. This operation was repeated (3.times.).
Hydrogenation was continued 18 hours. TLC indicated that a large amount of
starting material remained. The catalyst was removed by filtration, the
pad was washed with more ethanol and the filtrate was taken to dryness at
reduced pressure. The residue was dissolved in ethanol (100 mL), acetyl
chloride (1.5 mL) and 10% palladium on carbon (300 mg) were added and the
mixture was hydrogenated on the Parr apparatus at 55 psi for twenty hours.
The catalyst was removed by filtration. The pad was washed with more
ethanol. The filtrate was concentrated in vacuo to obtain title amine
which was used without purification.
##STR186##
The crude Part E amine was dissolved in dimethylformamide (2.5 mL).
H-pyrazole-1-carboxamidine (263 mg, 1.8 mmol) and diisopropylethyl amine
(700 .mu.L) were added. The mixture was stirred 24 hours at room
temperature. Ether (15 mL) was then added. Gummy material precipitated.
The ether was decanted and the precipitate was washed with more ether. The
gummy material was dissolved in methanol and taken to dryness in vacuo.
The remaining material was purified by preparative HPLC (YMC S-15 ODS
50.times.500 mm column, eluting with 40% methanol in water, containing
0.1% TFA). Fractions containing clean title compound were combined and
lyophilized to provide a white solid which was used to obtain NMR spectra,
recovered, dissolved in water (30 mL) and relyophilized to give title
compound (271 mg, 34%).
.alpha.!.sub.D =+39.7.degree. (c=0.6, MeOH)
Analysis: Calcd for 1.20 TFA+0.3 H.sub.2 O: C, 47.20; H, 5.81; N, 13.54; F,
11.02; S, 5.16. Found: C, 47.26; H, 5.86; N, 13.48; F, 10.93; S, 5.24.
EXAMPLE 34a
##STR187##
A solution of 1-BOC-2-(methylazido)pyrrolidine (1.36 g, 6.0 mmol) in
dichloromethane (5 mL) and trifluoroacetic acid (4 mL) was stirred at room
temperature for two hours. The reaction mixture was concentrated under
reduced pressure and toluene was added and removed at reduced pressure.
This process was repeated two more times. The crude amine,
N-BOC-L-aspartic acid, methyl ester (1.48 g, 6 mmol) and
1-hydroxybenzotriazol (1.01 g, 7.5 mmol) were dissolved in DMF (30 mL) and
treated with 4-methyl morpholine (1.32 mL, 12 mmol) followed by WSC (1.2
g, 6.0 mmol). The reaction mixture was stirred overnight at room
temperature. The mixture was diluted with ethyl acetate (70 mL) and washed
with satd. KHSO.sub.4 solution (20 mL), NaHCO.sub.3 solution (25 mL) and
10% lithium chloride solution (2.times.15 mL), dried over magnesium
sulfate and concentrated in vacuo to give title compound as a foam (1.68
g, 84%) which was used without purification.
##STR188##
A solution of Part A compound (1.68 g, 5.03 mmol) in dichloromethane (5 mL)
and trifluoroacetic acid (4 mL) was stirred at room temperature for two
hours. The reaction mixture was concentrated under reduced pressure and
toluene was added and removed at reduced pressure. This process was
repeated two more times. A solution of the crude des-BOC-amine and
napthalee-2-sulfonyl chloride (1.25 g, 5.5 mmol) in dichloromethane (25
mL) was cooled to 0.degree.-5.degree. C. and triethylamine (2.1 mL, 15
mmol) was added dropwise. The mixture was stirred cold for two hours and
then diluted with 1N HCl solution (30 mL). The dichloromethane layer was
separated, dried over magnesium sulfate and concentrated in vacuo. The
crude oil was chromatographed on silica gel, eluting with 50% and 75%
ethyl acetate in hexanes to give the title sulfonamido adduct as a white
foam (1.53 g, 68%).
##STR189##
The Part B azide (1.5 g, 3.37 mmol) was dissolved in absolute ethanol (80
mL) and treated with 10% palladium on carbon (300 mg). The reaction flask
was equipped with a hydrogen filled balloon via a three-way stopcock. Air
inside the flask was evacuated under reduced pressure and replaced with
hydrogen from the balloon. This operation was repeated (3.times.).
Hydrogenolysis was continued for twenty hours. The balloon was removed and
the catalyst was removed by filtration. The pad was washed with more
ethanol. The filtrate was concentrated in vacuo to obtain title amine
(Quant.) as a foam which was used without purification.
##STR190##
Part C amine (3.37 mmol) and N-carbobenzyloxy-3(S)-piperidine carboxylic
acid (790 mg, 3.7 mmol) were dissolved in DMF (20 mL) at RT. HOBT (500 mg,
3.7 mmol), 4-methyl morpholine (1.5 mL) and WSC (708 mg, 3.7 mmol) were
added. The reaction was stirred 20 hours at room temperature. The mixture
was then diluted with ethyl acetate (60 mL) and washed with KHSO.sub.4
solution (20 mL), saturated NaHCO.sub.3 solution (15 mL) and 10% lithium
chloride solution (2.times.15 mL), dried over magnesium sulfate and
concentrated in vacuo. The material obtained was chromatographed on silica
gel, eluting with ethyl acetate followed by 5% methanol in ethyl acetate
to give title compound (1.56 g, 72%) as a white foam.
##STR191##
Part D compound (625 mg, 0.97 mmol) was dissolved in methanol (60 mL) and
treated with Pearlman's catalyst (200 mg). The reaction flask was
connected to a hydrogen filled balloon via a threeway stopcock. Air inside
the flask was evacuated under reduced pressure and replaced with hydrogen
from the balloon. This operation was repeated (3.times.). Hydrogenation
was continued 18 hours. TLC indicated that the reaction was complete. The
catalyst was removed by filtration, the pad was washed with more methanol
and the filtrate was taken to dryness at reduced pressure to give title
amine (470 mg, 96%) which was used without purification.
##STR192##
The crude Part E amine (470 mg, 0.928 mmol) was dissolved in
dimethylformamide (2.0 mL). H-pyrazole-1-carboxamidine (190 mg, 1.3 mmol)
and diisopropylethylamine (360 .mu.L, 2 mmol) were added. The mixture was
stirred over a weekend at room temperature. Ether (25 mL) was then added.
Gummy material precipitated. The ether was decanted and the precipitate
was washed with more ether. The gummy material was dissolved in methanol
and taken to dryness in vacuo. The remaining material was purified by
preparative HPLC (YMC S-15 ODS 50.times.500 mm column, eluting with 53%
methanol in water, containing 0.1% TFA). Fractions containing clean title
compound were combined and lyophilized to provide a white solid which was
used to obtain NMR spectra, recovered, dissolved in water (30 mL) and
relyophilized to give title compound (324 mg, 49%).
.alpha.!.sub.D =-20.3.degree. (c=0.6, MeOH)
Analysis: Calcd for 1.10 TFA+0.9 H.sub.2 O: C, 49.10; H, 5.49; N, 11.77; F,
8.78; S, 4.49. Found: C, 49.11; H, 5.23; N, 11.58; F, 6.96; S, 4.42.
EXAMPLE 35a
1S(2R*,3R*)!-N-2-1-(Aminoiminomethyl)-3-piperidinyl!carbonyl!amino!me
thyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-.alpha.-toluenesulfonam
ide
##STR193##
Example 30a Part G(1) BOC-azide (3.85 mg, 9.55 mmol) was dissolved in TFA
(20 mL) and stirred at RT 1.5 hours. The TFA was removed by distillation
under reduced pressure and by coevaporation with toluene. The residue is
dissolved in dichloro-methane (100 mL), cooled in an ice bath, and
triethylamine (4.0 mL, 28.7 mmol) and .alpha.-toluene sulfonyl chloride
(2.00 g, 10.5 mmol) are added. The cooling bath is removed and the
solution is stirred for 2 h. This solution is washed with KHSO.sub.4
solution (25 mL.times.2) and NaHCO.sub.3 solution(25 mL.times.2), dried
over magnesium sulfate and evaporated to provide crude title compound. The
crude product is chromatographed on silica gel and eluted with 30% and 50%
EtOAc in hexane to provide title sulfonamide (2.597 g, 55%).
B.
1S(2R*,3R*)!-N-2-1-(Aminoiminomethyl)-3-piperidinyl!carbonyl!amino!m
ethyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-.alpha.-toluenesulfoma
nide
The title compound is prepared from Part A azide following the procedure
described in Example 26a.
EXAMPLE 36a
1S(2R*,3R*)!-N-2-1-(Aminoiminomethyl)-3-piperidinyl!carbonyl!amino!me
thyl!-1-pyrrolidinyl!-1-(carbomethoxymethyl)-2-oxoethyl!-.alpha.-t-toluenes
ulfonamide
##STR194##
A solution of Example 34a Part A compound (1.68 g, 5.03 mmol) in
dichloromethane (5 mL) and trifluoroacetic acid (4 mL) was stirred at room
temperature for two hours. The reaction mixture was concentrated under
reduced pressure and toluene was added and removed at reduced pressure.
This process was repeated two more times. A solution of the crude
des-BOC-amine and .alpha.-toluenesulfonyl chloride (1.14 g, 6 mmol) in
dichloromethane (25 mL) is cooled to 0.degree.-5.degree. C. and
triethylamine (2.1 mL, 15 mmol) is added dropwise. The mixture is stirred
cold for two hours and then diluted with 1N HCl solution (30 mL). The
dichloromethane layer is separated, dried over magnesium sulfate and
concentrated in vacuo. The crude oil is chromatographed on silica gel,
eluting with 50% and 75% ethyl acetate in hexanes to give the title
sulfonamido adduct (1.53 g, 74%).
B.
1S(2R*,3R*)!-N-2-1-(Aminoiminomethyl)-3-piperidinyl!carbonyl!amino!m
ethyl!-1-pyrrolidinyl!-1-(carbomethoxymethyl)-2-oxoethyl!-.alpha.-toluenesu
lfonamide
The title compound is prepared from Part A compound, following the
procedure described in Example 34a.
EXAMPLE 37a
1S(2R*,3R*)!-N-2-1-(Aminoiminomethyl)-3-piperidinyl!carbonyl!amino!met
hyl!-1-pyrrolidinyl!-1-(carboxamidomethyl)-2-oxoethyl!-.alpha.-toluenesulfo
namide
##STR195##
A solution of Example 36a Part A compound (816 mg, 2 mmol) in a saturated
methanolic ammonia (5 mL) is stirred at room temperature in a sealed tube
for 48 h. The mixture is concentrated under reduced pressure and in vacuo
to form the title product.
B.
1S(2R*,3R*)!-N-2-1-(Aminoiminomethyl)-3-piperidinyl!carbonyl!amino!m
ethyl!-1-pyrrolidinyl!-1-(carboxamidomethyl)-2-oxoethyl!-.alpha.-toluenesul
fonamide
The title compound is prepared from Part A compound, following the
procedure described in Example 34a.
EXAMPLES 38a TO 54a
The following compounds were prepared carrying out procedures described in
the specification and working examples.
38a.
S-(R*,R*)!-3-(Aminomethyl)-N-1-3-hydroxy-2-(2-napthalenylsulfonyl)ami
no!-1-oxopropyl!-2-pyrrolidinyl!methyl!benzamide, trifluoroacetate (1:1)
salt
.alpha.!.sub.D =-39.4+ (c 0.50, MeOH)
Anal. Calc'd for C.sub.26 H.sub.30 N.sub.4 O.sub.5 S.1.26 TFA.0.97 H.sub.2
O: C, 50.99; H, 4.98; N, 8.34; S, 4.77; F, 10.69 Found: C, 50.99; H, 4.83;
N, 8.35; S, 4.57; F, 10.72
39a.
S-(R*,R*)!-4-(Aminomethyl)-N-1-3-hydroxy-2-(2-napthalenylsulfonyl)ami
no!-1-oxopropyl!-2-pyrrolidinyl!methyl!benzeneacetamide, trifluoroacetate
(1:1) salt
.alpha.!.sub.D =-43.0.degree. (c 0.50, MeOH)
Anal. Calc'd for C.sub.27 H.sub.32 N.sub.4 O.sub.5 S.1.42 TFA.0.74 H.sub.2
O: C, 51.21; H, 5.03; N, 8.01; S, 4.58; F, 11.56 Found: C, 51.21; H, 4.95;
N, 8.08; S, 4.31; F, 11.56
40a.
S-(R*,R*)!-3-(Aminomethyl)-N-1-3-hydroxy-2-(2-napthalenylsulfonyl)ami
no!-1-oxopropyl!-2-pyrrolidinyl!METHYL!benzeneacetamide, trifluoroacetate
(1:1) salt
.alpha.!.sub.D =-44.1.degree. (c 1.00, MeOH)
Anal. Calc'd for C.sub.27 H.sub.33 N.sub.4 O.sub.5 S.1.35 TFA.1.48 H.sub.2
O: C, 50.58; H, 5.19; N, 7.94; S, 4.55; F, 10.91 Found: C, 50.58; H, 4.99;
N, 7.84; S, 4.47; F, 11.09
41a.
3S-3R*,3(R*,R*)!!-l-(Aminoiminomethyl)-N-1-2-(2-napthalenylsulfonyl)
amino!-1-oxopropyl!-2-pyrrolidinyl!METHYL!-3-piperidinecarboxamide,
trifluoroacetate (2:3) salt
.alpha.!.sub.D =-18.3.degree. (c 0.52, MeOH)
Anal. Calc'd for C.sub.25 H.sub.34 N.sub.6 O.sub.4 S.1.67 TFA.0.23 H.sub.2
O: C, 48.00; H, 5.13; N, 11.85; S, 4.52; F, 13.42 Found: C, 48.00; H,
5.37; N, 11.94; S, 4.45; F, 13.43
42a.
3S-3R*,3(R*,R*)!!-1-(Aminoiminomethyl)-N-1-3-hydroxy-2-(4-methylphe
nyl)sulfonyl!amino!-1-oxopropyl!-2-pyrrolidinyl!methyl!-3-piperidinecarboxa
mide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =+16.0.degree. (c 0.50, MeOH)
Anal. Calc'd for C.sub.22 H.sub.34 N.sub.6 O.sub.5 S.1.1 TFA.1.32 H.sub.2
O: C, 45.15; H, 5.91; N, 13.05; S, 4.98; F, 9.74 Found: C, 45.15; H, 5.52;
N, 12.88; S, 4.95; F, 9.53
43a.
3S-3R*,3(R*,R*)!!-1-(Aminoiminomethyl)-N-1-3-hydroxy-2-carbobenzylo
xy!amino!-1-oxopropyl!-2-pyrrolidinyl!methyl!-3-piperidinecarboxamide,
trifluoroacetate (1:1) salt
Anal. Calc'd for C.sub.23 H.sub.34 N.sub.6 O.sub.5 S.1.05 TFA.1.60 H.sub.2
O: C, 48.38; H, 6.19; N, 13.49; F, 9.60 Found: C, 48.39; H, 5.85; N,
13.16; F, 9.45
44a.
1S2(R*,S*)!!-N-2-1-(aminoiminomethyl)-4-piperidinyl!carbonyl!amino!
methyl!-1-piperidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonami
de, trifluoroacetate (1:1) salt
.alpha.!.sub.D =+0.55.degree. (c=0.5, MeOH), Analysis: Calcd for 0.95
TFA+1.7 H.sub.2 O: C, 49.02; H, 5.95; N, 12.29; F, 7.92; S, 4.69. Found:
C, 49.15; H, 5.61; N, 11.95; F, 7.68; S, 4.89.
45a.
S-(R*,R*)!-3-(aminoiminomethyl)amino!-N-1-3-hydroxy-2-2-naphthalene
sulfonyl!-amino!-1-oxopropyl!-2-pyrrolidinyl!methyl!propionamide,
trifluoroacetate (1:1) salt
.alpha.!.sub.D =-36.8.degree. (c=0.6, MeOH),
Analysis: Calcd for 1.20 TFA+0.50 H.sub.2 O: C, 46.05; H, 5.10; N, 13.21;
F, 10.75; S, 5.04. Found: C, 46.05; H, 4.98;, N, 13.02; F, 10.82; S, 4.97.
46a.
1S2(R*,S*),(3R*)!!-N-2-1-(aminoiminomethyl)-3-piperidinyl!carbonyl!
amino!methyl!-1-piperidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesul
fonamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =+24.1.degree. (c=0.9, MeOH),
Analysis: Calcd for 1.20 TFA+0.7 H.sub.2 O: C, 49.14; H, 5.61; N, 12.11; F,
9.85; S, 4.62. Found: C, 49.17; H, 5.36; N, 12.04; F, 9.70; S, 4.70.
47a.
1S2R*!!-N-2-1-(aminoiminomethyl)-3-phenyl!carbonyl!amino!methyl!-1-
pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenesulfonamide,
trifluoroacetate (1:1) salt
.alpha.!.sub.D =-41.0.degree. (c=0.4, MeOH),
Analysis: Calcd for 1.20 TFA+1.1 H.sub.2 O: C, 50.14 H, 4.80; N, 10.30; F,
10.05; S, 4.71. Found: C, 50.13; H, 4.52;, N, 10.08; F, 9.78; S, 4.42.
48a.
1S2R*!!-N-2-1-(aminoiminomethyl)-cis-5-methyl-3-piperidinyl!carbonyl
!amino!methyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthalenes
ulfonamide, trifluoroacetate(1:1) salt
.alpha.!D=-25.5.degree. (c=0.4, MeOH)
Analysis: Calcd for 1.25 TFA+1.2 H.sub.2 O: C, 48.29; H, 5.64; N, 11.86; F,
10.05; S, 4.52 Found: C, 48.34; H, 5.54; N, 11.91; F, 9.99; S, 4.55.
49a.
1S2R*!!-N-2-1-(aminoiminomethyl)-trans-5-methyl-3-piperidinyl!carbo
nyl!amino!methyl!-1-pyrrolidinyl!-1-(hydroxymethyl)-2-oxoethyl!-2-naphthale
nesulfonamide, trifluoroacetate(1:1) salt
.alpha.!.sub.D =-28.8.degree. (c=0.6, MeOH),
Analysis: Calcd for 1.10 TFA+1.0 H.sub.2 O: C, 49.22; H, 5.73; N, 12.21; F,
9.11; S, 4.66. Found: C, 48.23; H, 5.66; N, 12.08; F, 9.12; S, 4.87.
50a.
1S(2R*,3R*,4R*)!-N-2-1-(aminoiminomethyl)-3-piperidinyl!carbonyl!ami
no!methyl!-1-pyrrolidinyl!-1-(1-hydroxyethyl)-2-oxoethyl!-2-naphthalenesulf
onamide, trifluoroacetate (1:1) salt
.alpha.!D=-10.0.degree. (c=0.7, MeOH),
Analysis: Calcd for 1.10 TFA+1.7 H.sub.2 O: C, 48.34; H, 5.83; N, 11.99; F,
8.95; S, 4.58. Found: C, 48.32; H, 5.57; N, 11.84; F, 8.83; S, 4.71.
51a.
1S(2R*,3S*,4R*)!-N-2-1-(aminoiminomethyl)-3-piperidinyl!carbonyl!ami
no!methyl!-1-pyrrolidinyl!-1-(1-hydroxyethyl)-2-oxoethyl!-2-naphthalenesulf
onamide, trifluoroacetate (1:1) salt
.alpha.!.sub.D =-19.5.degree. (c=0.6, MeOH),
Analysis: Calcd for 1.10 TFA+1.0 H.sub.2 O: C, 49.22; H, 5.73; N, 12.21; F,
9.11; S, 4.66. Found: C, 49.12; H, 5.71; N, 12.16; F, 9.38; S, 4.99.
52a.
R-(S*,R*)!-N-2-2-1-(aminoiminomethyl)-4-piperidinyl!carbonyl!amino!
methyl!-1-pyrrolidinyl!-1-(phenylmethyl)-2-oxoethyl!methanesulfonamide,
trifluoroacetate (1:1) salt
.alpha.!.sub.D =-34.7.degree. (c=0.6, MeOH),
Analysis: Calcd for 1.25 TFA+0.2 H.sub.2 O: C, 47.10; H, 5.75; N, 13.45; F,
11.40; S, 5.13. Found: C, 47.09; H, 5.84; N, 13.15; F, 11.44; S, 5.32.
53a.
S-(R*,R*)!-N-2-1-(aminoiminomethyl)-3-piperidinyl!carbonyl!amino!met
hyl!-1-pyrrolidinyl!-2-oxoethyl!-2-naphthalenesulfonamide, trifluoroacetate
(1:1) salt
.alpha.!.sub.D =+9.1.degree. (c=0.3, MeOH),
Analysis: Calcd for 1.20 TFA +0.8 H.sub.2 O: C, 48.64; H, 5.38; N, 12.89;
F, 10.49; S, 4.92. Found: C, 48.95; H, 5.23; N, 12.39; F, 10.32; S, 4.93.
54a
R-(S*,R*)!-N-2-2-1-(aminoiminomethyl)-4-piperidinyl!carbonyl!amino!
methyl!-1-pyrrolidinyl!-1-(methyl)-2-oxoethyl!benzylsulfonamide,
trifluoroacetate (1:1) salt
.alpha.!.sub.D =+13.5.degree. (c=0.7, MeOH),
Analysis: Calcd for 1.30 TFA+0.2 H.sub.2 O: C, 46.87; H, 5.71; N, 13.33; F,
11.75; S, 5.09. Found: C, 47.23; H, 5.78; N, 12.86; F, 11.36; S, 5.29.
The following Examples represent preferred embodiments of the fourth
embodiment of the present invention. Unless otherwise indicated, all
temperatures are expressed in degrees Centigrade.
EXAMPLE A
N-1-(Aminoiminomethyl)-4-piperidinyl!methyl!-1-D-phenylalanyl-L-prolinami
de
A.
4-(Aminomethyl)-N,N'-bis(1,1-dimethylethoxy)carbonyl!-1-piperidinecarboxi
midamide
To a stirred solution of 4-aminomethylpiperidine (0.72 g, 6.31 mmol) in 40
mL of toluene was added benzaldehyde (0.78 mL, 6.94 mmol). The reaction
solution was refluxed for 18 h and water was removed by a Dean Stark trap.
The reaction solution was cooled to room temperature at which time bis-Boc
amidinopyrazole (1.96 g, 6.31 mmol) was added. The reaction solution was
stirred at room temperature for 48 h and concentrated in vacuo. The oily
residue was diluted with 15 mL of 1M (aq) KHSO.sub.4 solution and stirred
at room temperature for 5 h. This aqueous solution was washed with ether
(2.times.20 mL) and basified to pH 12 by the addition of 1N NaOH solution.
This basic solution was then saturated with NaCl and extracted with
dichloromethane (3.times.60 mL). The combined dichloromethane extracts
were dried (MgSO.sub.4), filtered and concentrated in vacuo to give 2.10 g
(93%) of title amine which was used for the next transformation without
further purification.
B.
(S*)-N-1-(1,1-Dimethylethoxy)carbonyl!amino!(1,1-dimethylethoxy)car
bonyl!imino!methyl!-4-piperidinyl!-methyl!-1-1-(1,1-dimethylethoxy)carbo
nyl!amino!-2-phenylethyl!-S-prolinamide
To a stirred solution of N-Boc-D-Phe-L-Pro-OH (0.73 g, 2.02 mmol), Part A
amine (0.72 g, 2.02 mmol) and 1-hydroxybenzotriazole monohydrate (0.34 g,
2.02 mmol) in 30 mL of DMF was added in order 4-methylmorpholine (0.66 mL,
6.05 mmol) and ethyl-3-(3-dimethylamino)propyl carbodiimide hydrochloride
(0.39 g, 2.02 mmol). The reaction solution was stirred at room temperature
for 19 h and concentrated under pump vacuum at 45.degree. C. The residue
was diluted with 100 mL of saturated NaHCO.sub.3 solution and extracted
with dichloromethane (4.times.100 mL). The combined dichloromethane
extracts were dried (MgSO.sub.4), filtered and concentrated in vacuo. This
was chromatographed on silica gel to give 0.70 g (50%) of title bis-Boc
guanidine.
C.
N-1-(Aminoiminomethyl)-4-piperidinyl!-methyl!-1-D-phenylalanyl-L-prolina
mide
To a stirred solution of Part B bis-Boc guanidine (0.68 g, 0.97 mmol) in
6.0 mL of dichloromethane was added trifluoroacetic acid (TFA) (6.00 mL,
77.9 mmol). The reaction solution was stirred at room temperature for 3 h
and concentrated in vacuo. This was purified by prep HPLC to give 310 mg
(45%) of title compound.
EXAMPLE B
N-1-(Aminoiminomethyl)-3-piperidinyl!methyl!-1-D-phenylalanyl-L-prolinami
de
5A. N-Boc-3-hydroxymethylpiperidine
To a stirred solution of 3-hydroxymethylpiperidine (15.1 g, 131 mmol) and
Et.sub.3 N (21.9 mL, 158 mmol) in 100 mL of dichloromethane was added
dropwise a solution of di-t-butyl dicarbonate (31.5 g, 144 mmol) in 100 mL
of dichloromethane over 1 h. The reaction was stirred at room temperature
for 18 h and then diluted with 200 mL of dichloromethane. The resulting
solution was washed with 1N HCl solution (3.times.100 mL), saturated
NaHCO.sub.3 solution (2.times.100 mL) and brine (1.times.100 mL). The
organic layer was dried (MgSO.sub.4), filtered and concentrated in vacuo
to give N-Boc-3-hydroxymethylpiperidine (27.0 g, 96%).
B. 3-(Azidomethyl)-1-piperidinecarboxylic acid, 1,1-dimethylethyl ester
To a stirred solution of Part A N-Boc-3-hydroxymethylpiperidine (27.0 g,
126 mmol) in 150 mL of dry dichloromethane under argon at 0.degree. C. was
added in order triethylamine (22.7 mL, 163 mmol) and methanesulfonyl
chloride (11.7 mL, 151 mmol). The reaction was stirred at room temperature
for 1.5 h and diluted with 450 mL of dichloromethane. The reaction was
washed with 0.degree. C. 1N HCl solution (2.times.100 mL) and brine
(1.times.100 mL). The dichloromethane layer was dried (Na.sub.2 SO.sub.4),
filtered and concentrated in vacuo. The residue was dissolved in 200 mL of
DMF and combined with sodium azide (24.5 g, 377 mmol). The mixture was
stirred at room temperature for 33 h and the solid was filtered off. The
filtrate was concentrated under pump vacuum at 45.degree. C. The residue
was partitioned between 400 mL of EtOAc and 10% sodium thiosulfate
solution (2.times.100 mL) and brine (1.times.100 mL). The EtOAc layer was
dried (MgSO.sub.4), filtered and concentrated in vacuo. Purification was
effected by a flash column chromatography on silica gel to give 19.5 g
(65%) of title azide.
C. 3-(Aminomethyl)-1-piperidinecarboxylic acid, 1,1-dimethylethyl ester
To a stirred solution of Part B azide (19.0 g, 79.2 mmol) in 250 mL of
methanol under argon was added 10% Pd/C (3.80 g, 20% based on the weight
of Part B azide). The atmosphere was replaced with hydrogen by several
vacuum-fill cycles. The mixture was stirred at room temperature for 15 h.
The catalyst was filtered through a 4 .mu.M polycarbonate film and rinsed
with methanol (4.times.30 mL). The filtrate was concentrated in vacuo to
give 16.3 g (96%) of title amine.
D.
N-1-(1,1-Dimethylethory)carbonyl!-3-piperidinyl!methyl!-1-N-(phenylme
thoxy)carbonyl!-D-phenylalanyl!-L-prolinamide
To a stirred solution of Part C amine (2.00 g, 9,35 mmol),
N-Cbz-D-Phe-L-Pro (3.70 g, 9.35 mmol), 1-hydroxybenzotriazole monohydrate
(1.58 g, 9.35 mmol) and 4-methylmorpholine (3.07 mL, 28.0 mmol) was added
ethyl-3-(3-dimethylamino)propyl carbodiimide hydrochloride (1.79 g, 9.35
mmol). The reaction solution was stirred at room temperature for 17 h and
concentrated under pump vacuum at 45.degree. C. The residue was dissolved
in 360 mL of EtOAc and washed with 1N HCl solution (2.times.120 mL),
saturated NaHCO.sub.3 solution (1.times.120 mL) and brine (1.times.120
mL). The EtOAc layer was dried (MgSO.sub.4), filtered, concentrated in
vacuo and chromatographed on silica gel to give 1.30 g (23%) of title
carbamate.
E.
N-1-(Aminoiminomethyl)-3-piperidinyl!methyl!-1-N-(phenylmethoxy)carbon
yl!-D-phenylalanyl!-L-prolinamide
To a stirred solution of Part D carbamate (2.30 g, 3.89 mmol) in 10 mL of
dry dichloromethane was added 0.degree. C. 4N HCl in dioxane (15.0 mL,
60.0 mmol). The solution was stirred at room temperature for 3 h and
diluted with 300 mL of ether. The precipitate was filtered off and rinsed
with ether (3.times.30 mL). The precipitate was dried under pump vacuum at
room temperature and purified by prep HPLC to give 1.39 g (59%) of
intermediate amine.TFA. To a stirred solution of the intermediate
amine.TFA salt (500 mg, 0.83 mmol) and diisopropylethyl amine (0.35 mL,
1.98 mmol) in 2.0 mL of DMF was added 1H-pyrazole-1-carboxamidine (133 mg,
0.91 mmol). The reaction solution was stirred at room temperature for 6 h
and diluted with 100 mL of ether. The desired oily precipitate was
separated from the ether solution and purified by prep HPLC to give 250 mg
(47%) of title Cbz-carbamate.
F.
N-1-(Aminoiminomethyl)-3-piperidinyl!methyl!-1-D-phenylalanyl-n-prolinam
ide
To a stirred solution of Part E Cbz-carbamate (240 mg, 0.37 mmol) in 10 mL
of methanol under argon was added 20% Pd(OH).sub.2 /C (48 mg, 20% based on
the weight of Part E Cbz carbamate). The atmosphere was replaced with
hydrogen by several vacuum-fill cycles. The reaction mixture was stirred
at room temperature for 24 h. The catalyst was filtered off and rinsed
with methanol (4.times.20 mL). The filtrate was concentrated in vacuo. The
residue was dissolved in 50 mL of a solution of 0.1% TFA in water and
lyophilized to give 220 mg (82%) of title compound.
Following the procedures of Examples a. and b., the following examples of
compounds of the invention may be prepared.
TABLE
__________________________________________________________________________
##STR196##
Example No.
R.sup.3 R R.sup.2
R.sup.1
n p o A R.sup.4
__________________________________________________________________________
c CBZ CH.sub.2 OH(S)
H CH.sub.3
1 0 --
##STR197
#
##STR198##
d H H OH H 0 1 CO
##STR199
#
##STR200##
e C.sub.6 H.sub.5 CO
CH.sub.2 C.sub.6 H.sub.5 (R)
OCH.sub.3
CH.sub.3
0 2 --
##STR201
#
##STR202##
e CO.sub.2 CH.sub.3
CH.sub.2 C.sub.6 H.sub.5 (S)
CH.sub.3
CH.sub.3
1 1 --
##STR203
#
##STR204##
f CO.sub.2 C.sub.6 H.sub.5
CH.sub.2 CH.sub.2 CONH.sub.2 (S)
H H 1 0 --
##STR205##
CH.sub.2 NH.sub.2
g CH.sub.3 CO
CH.sub.2 CH.sub.2 CONH.sub.2 (R)
H CH.sub.3
1 1 CO
##STR206##
CH.sub.2 NH.sub.2
##STR207##
CH(OH)CH.sub.3 (S-Thr)
CHCH.sub.2 CH.sub.2 CH
0 2 --
##STR208
#
##STR209##
i H CH(OH)CH.sub.3 (S-alloThr)
CH.sub.3
H 1 2 CO
##STR210
#
##STR211##
j C.sub.6 H.sub.5 CH.sub.2 CO
##STR212## SCH.sub.3
CH.sub.3
1 1 --
##STR213
#
##STR214##
k CO.sub.2 C.sub.6 H.sub.5
CH.sub.2 CH.sub.2 CO.sub.2 H(R)
H CH.sub.3
0 0 CO
##STR215
#
##STR216##
l CO.sub.2 CH.sub.2 C.sub.6 H.sub.5
CH.sub.2 OCH.sub.2 Ph(R)
H H 1 0 --
##STR217
#
##STR218##
m H CH.sub.2 CH.sub.2 Ph(S)
H H 1 1 --
##STR219
#
##STR220##
n H
##STR221## H H 1 2 --
##STR222
#
##STR223##
__________________________________________________________________________
EXAMPLE (I)
N-4-(Aminoiminomethyl)amino!butyl!-1-N-(methylsulfonyl)-D-phenylalanyl!-
L-prolinamide, trifluoroacetate (1:1) salt
##STR224##
To a stirred solution of 1,4-diaminobutane (50 g, 567 mmol) in 195 mL of
dioxane under argon at room temperature was added dropwise a solution of
di-t-butyl dicarbonate (15.7 g, 71.9 mmol) in 195 mL of dioxane over 3.5
h. Some white precipitate appeared during the addition. The mixture was
stirred at room temperature for 22 h and concentrated in vacuo. The
residue was diluted with 320 mL of water and the precipitate was filtered
off. The aqueous filtrate was extracted with methylene chloride
(3.times.300 mL). The combined methylene chloride extracts were washed in
water (2.times.200 mL) and brine (1.times.200 mL). The organic layer was
dried (MgSO.sub.4), filtered and concentrated in vacuo to give 9.79 g
(72%) of title mono-BOC.amine.
##STR225##
To a stirred solution of N-CBz-L-proline (12.7 g, 50.9 mmol),
1-hydroxybenzotriazole monohydrate (6.49 g, 50.9 mmol) and Part A
BOC.amine (9.57 g, 50.9 mmol) in 250 mL of DMF was added in order
4-methylmorpholine (11.2 mL, 102 mmol) and ethyl-3-(dimethylamino)propyl
carbodiimide hydrochloride (9.76 g, 50.9 mmol). The reaction solution was
stirred at room temperature for 22 h and concentrated under pump vacuum at
50.degree. C. The residue was diluted with 600 mL of EtOAc and washed with
1N HCl solution 2.times.250 mL), saturated NaHCO.sub.3 solution
(2.times.250 mL) and brine (1.times.250 mL). The EtOAc layer was dried
(MgSO.sub.4), filtered and concentrated in vacuo to give 20.7 g 97%) of
title CBz.amine.
##STR226##
To a stirred solution of Part B CBz.amine (20.2 g, 48.2 mmol) in 250 mL of
methanol under argon was added 20% Pd(OH).sub.2 /C (4.04 g, 20% based on
the weight of Part B amine). The atmosphere was replaced by hydrogen with
several vacuum-fill cycles. The reaction mixture was stirred at room
temperature for 21 h. The catalyst was filtered off through a 4 .mu.M
polycarbonate film and rinsed with methanol (3.times.50 mL). The filtrate
was concentrated in vacuo. The oily residue was dissolved in 200 mL of
ether and treated with 1N HCl solution in ether (53.0 mL, 53.0 mmol). The
solution was concentrated in vacuo. The residue was mixed with 300 mL of
toluene and 30 mL of methanol and concentrated in vacuo to give title
amine hydrochloride in a quantitative yield (15.5 g) as an oil.
##STR227##
A stirred solution of N-.alpha.-CBZ-D-phenylalanine (0.56 g, 1.9 mmol) in
6.5 mL of DMF at room temperature under argon was treated with
1-hydroxybenzotriazole (0.29 g, 1.9 mmol) and EDAC* (0.36 g, 1.9 mmol).
After 20 minutes, Part C compound was added (0.50 g, 1.6 mmol) and
stirring was carried out for 16 hours. The reaction was quenched by the
addition of 75 mL of 0.25M KHSO.sub.4 solution. The suspension was washed
with EtOAc (2.times.40 mL), the combined EtOAc layers were washed with
0.25M KHSO.sub.4 solution (2.times.40 mL), saturated aqueous KHCO.sub.3
solution (2.times.40 mL), brine, dried (Na.sub.2 SO.sub.4), and
concentrated to yield 1.07 g of a white taffy, which by TLC analysis
appeared to contain unreacted N-.alpha.-CBZ-D-phenylalanine. The crude
product was redissolved in 60 mL of EtOAc, washed with saturated aqueous
KHCO.sub.3 solution (3.times.40 mL), brine, dried (Na.sub.2 SO.sub.4),
concentrated, co-evaporated several times with ether and hexane and
triturated with 50 mL of hexane to yield title compound (0.78 g, 88%) as a
colorless solid.
##STR228##
Trifluoroacetic acid (3.2 mL) was added to ice-cooled Part D compound (0.78
g, 1.4 mmol). The reaction solution was stirred at room temperature for 2
hours and 45 minutes. Trifluoroacetic acid was removed under vacuum and
co-evaporated several times with ether and hexane to obtain. a colorless
taffy.
##STR229##
A solution of Part E compound (0.80 g, 1.38 mmol) in 10.9 mL of absolute
ethanol was treated with amidine sulfonic acid (0.26 g, 2.1 mmol) followed
by triethylamine (0.58 mL, 4.1 mmol). After addition of the triethylamine,
a yellow, homogeneous reaction solution slowly formed. After 2 hours, TLC
analysis of the reaction mixture indicated it was complete. The reaction
mixture was concentrated, dissolved in 25 mL of CH.sub.3 OH, and filtered.
Preparative HPLC of the filtered solution provided the title compound as a
colorless solid, (414.0 mg, 45%) mp 50.degree.-140.degree. C. with
foaming.
##STR230##
A solution of 0.20 g (0.30 mmol) of Part F compound in 1.5 mL of CH.sub.3
OH with 40 mg of Pearlman's catalyst was hydrogenated at 1 atm for 3
hours. The reduction was judged complete by TLC analysis after 2.5 hours.
The catalyst was removed by filtration and the filtrate was concentrated
to yield an oil, which was redissolved in 10 mL of CH.sub.3 OH, acidified.
with 0.20 mL of trifluoroacetic acid, concentrated, dissolved in H.sub.2 O
and lyophilized to yield title compound as a colorless solid (152.1 mg,
81%), mp 124.degree.-125.degree. C.
H.
N-4-(Aminoiminomethyl)amino!butyl!-1-N-(methylsulfonyl)-D-phenylalanyl!
-L-prolinamide, trifluoroacetate (1:1) salt
##STR231##
To a stirred solution of Part H compound (550 mg, 0.80 mmol) in 15 mL of
dry CH.sub.2 Cl.sub.2 and 15 mL of dry THF under argon was added Et.sub.3
N (0.44 mL, 3.20 mmol) followed by methanesulfonyl chloride (68.0 .mu.L,
0.88 mmol). The turbid mixture was stirred at room temperature for 3 h and
diluted with 0.50 mL of water. The mixture was stirred at room temperature
for 10 min and concentrated in vacuo, the residue diluted with 30 mL of
methanol and concentrated in vacuo. This material was purified by
preparative HPLC and lyophilized to give 300 mg (75%) of title compound.
Analysis for 1.15 CF.sub.3 COOH+0.75 H.sub.2 O: C, 44.85; H, 5.85; N,
14.07; S, 5.37; F, 10.97 Found: C, 45.02; H, 5.82; N, 13.94; S, 5.34; F,
10.89
Optical rotation: .alpha.!.sub.D =-73.2.degree. (c=1.00, MeOH)
Following the procedures of Example (i), the following examples of
compounds of the invention may be prepared.
TABLE
__________________________________________________________________________
##STR232##
Example No.
Alkyl.sub.1-2
R R.sup.2
R.sup.1
n m Y
__________________________________________________________________________
2(i) C.sub.2 H.sub.5
CH.sub.2 OH(S)
H H 1 2 NH
3(i) CH.sub.3
H OH H 0 1 NH
4(i) C.sub.2 H.sub.5
C.sub.2 C.sub.6 H.sub.5 (R)
OCH.sub.3
CH.sub.3
0 2 S
5(i) CH.sub.3
CH.sub.2 C.sub.6 H.sub.5 (S)
6(i) C.sub.2 H.sub.5
CH.sub.2 CH.sub.2 CONH.sub.2 (S)
H H 1 0 S
7(i) CH.sub.3
CH.sub.2 CH.sub.2 CONH.sub.2 (R)
H CH.sub.3
1 1 NH
8(i) C.sub.2 H.sub.5
CH(OH)CH.sub.3 (S-Thr)
CH.sub.2 CH.sub.2 CH.sub.2 CH
0 2 S
9(i) CH.sub.3
CH(OH)CH.sub.3 (S-alloThr)
CH.sub.3
H 1 2 NH
10(i) C.sub.2 H.sub.5
##STR233## SCH.sub.3
CH.sub.3
1 1 S
11(i) CH.sub.3
CH.sub.2 CH.sub.2 CO.sub.2 H(R)
H CH.sub.3
0 3 NH
12(i) C.sub.2 H.sub.5
CH.sub.2 OCH.sub.2 Ph(R)
H H 1 0 S
13(i) CH.sub.3
CH.sub.2 CH.sub.2 Ph(S)
H H 1 1 NH
__________________________________________________________________________
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